Claimed up to 83% Success Rate
Virginia Livingston, MD :
Cancer Quack Or Medical Genius?
By Alan Cantwell, MD
Cancer is the most frightening human disease and its cause remains
elusive. Therefore, it seems inconceivable that the discovery of a
germ cause of cancer would provoke such hostility among the cancer
establishment. But, in truth, the belief in a cancer germ has
always been the ultimate scientific heresy.
In the long history of cancer research there was never a physician
more outspoken and controversial than Virginia Wuerthele-Caspe
Livingston (1906-1990). For more than 40 years she championed the
revolutionary idea that bacteria caused cancer and devised a
treatment to try and combat these microbes by immunotherapy.
Sixteen years after her death she is now largely forgotten but
still condemned by such powerful organizations as the American
Cancer Society-and blacklisted on Quackwatch-a self-proclaimed
"non-profit corporation dedicated to combating health-related
frauds, myths, fads, and fallacies. "
LIVINGSTON'S CANCER RESEARCH
Beginning in the late 1940s, Livingston was able to grow bacteria
from cancer tumors; and when she and her associates injected
cancer bacteria into laboratory animals, some developed cancer.
Other animals developed degenerative and proliferative diseases,
and some animals remained healthy. Livingston believed the
"immunity" of the host was an important factor in determining
whether cancer would develop.
Virginia Livingston MD
In 1969 at a meeting at the New York Academy of Sciences ,
Livingston and her colleagues proposed that cancer was caused by a
highly unusual bacterium which she named Progenitor
cryptocides-Greek for 'ancestral hidden killer.' Neverthless,
Livingston claimed elements of the microbe were present in every
human cell. Due to its biochemical properties, she believed the
organism was responsible for initiating life and for the healing
of tissue-and for killing us with cancer and other infirmities.
Critics of this research continued to insist there was no such
thing as a cancer germ.
In her attempt to use a variety of modalities (diet, supplements,
antibiotics, as well as traditional methods) to treat cancer, she
utilized an 'autogenous' vaccine derived from the patient's own
cancer bacteria found in the urine and blood. Livingston explained
it was not an anti-cancer vaccine, but rather a vaccine to help
stimulate and improve the patient's own immune system. The
administration of this unapproved vaccine caused a furor in the
cancer establishment and eventually legal action was undertaken
against her and the Livingston-Wheeler Clinic in San Diego. In
spite of all her legal troubles, she continued seeing patients
until her death at 83.
In March 1990, the year of her death, a highly critical article on
the Livingston-Wheeler therapy appeared in the American Cancer
Society-sponsored CA: A Cancer Journal for Physicians. (No authors
were listed.) The report advised patients to stay away from the
San Diego clinic and claimed: "Livingston-Wheeler's cancer
treatment is based on the belief that cancer is caused by a
bacterium she has named Progenitor cryptocides. Careful research
using modern techniques, however, has shown that there is no such
organism and that Livingston-Wheeler has apparently mistaken
several different types of bacteria, both rare and common, for a
unique microbe. In spite of diligent research to isolate a
cancer-causing microorganism, none has been found. Similarly,
Livingston-Wheeler's autologous vaccine cannot be considered an
effective treatment for cancer. While many oncologists have
expressed the hope that someday a vaccine will be developed
against cancer, the cause(s) of cancer must be determined before
research can be directed toward developing a vaccine. The
rationale for other facets of the Livingston-Wheeler cancer
therapy is similarly faulty. No evidence supports her contention
that cancer results from a defective immune system, that a
whole-foods diet restores immune system deficiencies, that
abscisic acid slows tumor growth, or that cancer is transmitted to
humans by chickens." (The full report is on-line at:
BACTERIA AS A CAUSE OF CANCER
The recognition of disease-producing bacteria allowed medical
science to emerge from the dark ages into the era of modern
medicine. In the late nineteenth century when diseases like
tuberculosis (TB) , syphilis, and leprosy were proven to be caused
by bacteria, some doctors also suspected human cancer might have a
The idea that bacteria cause cancer is considered preposterous by
most physicians. However, despite the antagonistic view of the
American Cancer Society and medical science, there is ample
evidence in the published peer-reviewed literature that strongly
suggests that 'cancer microbes' cause cancer.
Intracellular variably-sized coccoid forms
in breast cancer.
Acid-fast stain; Magnification x1000, in oil
According to reports by Livingston and various other researchers,
cancer is caused by pleomorphic, cell wall deficient bacteria. The
various forms of the organism range in size from submicroscopic
virus-like forms, up to the size of bacteria, yeasts, and fungi.
In culture and in tissue the bacterial forms are variably
'acid-fast' (having a staining quality like TB bacteria). These
bacteria are ubiquitous and exist in the blood and tissues of all
human beings (yet another 'heresy'). In the absence of a
protective immune response, these cell wall deficient bacteria may
become pathogenic and foster the development of cancer ,
autoimmune disease, AIDS, and certain other chronic diseases of
Needless to say, all this research fell on dead ears because
bacteria were totally ruled out as the cause of all cancers in the
early years of the twentieth century. Thus, bacteria observed in
cancer were simply dismissed as elements of cellular degeneration,
or as invaders of tissue weakened by cancer, or as 'contaminants'
of laboratory origin.
LIVINGSTON AND PROGENITOR CRYPTOCIDES
Beginning in1950, in a series of papers and books, Livingston and
her co-workers claimed the cancer microbe was a great imitator
whose various pleomorphic forms resembled common staphylococci,
diphtheroids, fungi, viruses, and host cell inclusions. Yet if the
germ were studied carefully through all its transitional stages,
it could be identified as a single agent. She was the first to
suggest that the acid-fast stain was the key to the identification
of the cancer microbe in tissue and in culture; and also
demonstrated its appearance in the blood of cancer patients, by
use of dark-field microscopy.
Anyone who takes the time to read Livingston's reports in the
medical literature will quickly recognize that she was a credible
research scientist, who allied herself with other experts-and was
certainly not the quack doctor pictured by her detractors. Her
achievements in cancer microbiology can also be found in her
autobiographical books: Cancer, A New Breakthrough (1972); The
Microbiology of Cancer (1977); and The Conquest of Cancer (1984).
Her research has been confirmed by other scientists, such as
microbiologist Eleanor Alexander-Jackson, cell cytologist Irene
Diller, biochemist Florence Seibert, and dermatologist Alan
Cantwell, among others.
Intracellular bacteria in prostate cancer.
Acid-fast stain; magnification
x1000, in oil.
THE CANCER MICROBE AND BACTERIAL PLEOMORPHISM
Microbiologists have long resisted the idea of bacterial
pleomorphism, and do not recognize or accept the various growth
forms and the bacterial 'life cycle' proposed by various cancer
microbe workers. Most bacteriologists do not accept the idea of a
bacterium changing from a coccus to a rod, or to a fungus.
Depending on the environment, the microbe in its cell
wall-deficient phase may attain large size, even larger that a red
blood cell. Other forms are submicroscopic and virus-sized.
Electronic microscopic studies and photographs of filtered
(bacteria-free) cultures of the cancer microbe show virus-size
elements of the cancer microbe that can revert into
The cancer microbe has adapted to life in man and animals by
existing in a mycoplasma-like or cell wall deficient state. In
tissue sections of cancer stained for bacteria with the special
acid-fast stain, the microbe can be seen as a variably acid-fast
(blue, red, or purple-stained) round coccus or as barely visible
granules . At magnifications of one thousand times (in oil), these
forms can be observed within and also outside of the cells.
Careful study and observation of the tiny round coccoid forms in
cancer tissue indicate they can enlarge progressively up to the
size of so-called Russell bodies, which are well-known to
pathologists. Russell bodies can attain the size of red blood
cells, and even larger.
William Russell was a well-respected Scottish pathologist who in
1890 first reported the finding of 'cancer parasites' in the
tissue of all the cancers he studied. However, modern pathologists
deny that Russell's bodies are microbial in origin. For more
information on Russell bodies and Russell's 'cancer parasite' (and
its intimate relationship to cancer microbes), Google: The
forgotten clue to the bacterial cause of cancer; or go to:
OVERLOOKING HIDDEN BACTERIA IN CANCER
Once bacteria were eliminated as a cause of cancer a century ago,
it became dogma and impossible to change medical opinion. In this
current era of medical science, one would think it impossible for
infectious disease experts and pathologists to not recognize
bacteria in cancer. However, bacteria can still pop up in diseases
in which they were initially overlooked.
When a new and deadly lung disease broke out among legionnaires in
Philadelphia in July 1976, two hundred twenty-two people became
ill and thirty-four died. The cause of the killer lung disease
remained a medical mystery for over five months until Joe McDade
at the Leprosy Branch of the CDC detected unusual bacteria in
guinea pigs experimentally infected with lung tissue from the dead
legionnaires. Further modification of bacterial culture methods
finally allowed the isolation of the causative and previously
overlooked bacteria, now known as Legionella pneumophila.
Lymph node showing Hodgkin's lymphoma. Arrows point to
variably-sized round coccoid forms and larger Russell bodies. Gram
stain; magnification x1000, in oil.
Yet another example of dogma-defying research is provided by
recent studies proving that bacteria (Helicobacter pylori) are a
common cause of stomach ulcers, which can sometimes lead to
stomach cancer and lymphoma. For a century, physicians refused to
believe bacteria caused ulcers because they thought bacteria could
not live in the acid environment of the stomach. In 2005 the Nobel
Prize in Medicine was awarded to two Australian researchers for
their 1982 discovery. These stomach bacteria could only be
detected by use of special tissue stains. The CDC now claims that
H. pylori causes more than 90% of duodenal ulcers and 80% of
gastric ulcers. Approximately two-thirds of the world's population
is infected with these microbes.
In the past four years there have been medical reports of newly
discovered bacteria in serious lymph node disease; in Hodgkin's
lymphoma; in cancer of the mouth; and in prostate cancer, to name
only a few.
All these studies prove bacteria can pop up in diseases where they
are least expected. Such a caveat is appropriate for doctors who
think they know everything about cancer and who pooh-pooh all
aspects of cancer microbe research.
A CENTURY OF CANCER MICROBE RESEARCH
Livingston never claimed that she was the discoverer of the
microbe of cancer. In her writings she always gave credit to
various scientists, some dating back to the nineteenth century,
who attempted to prove that bacteria cause cancer. Some of these
remarkable researchers include the long-forgotten cancer microbe
studies of Scottish obstetrician James Young, Chicago physician
John Nuzum, Montana surgeon James Scott, the infamous psychiatrist
and cancer researcher Wilhelm Reich, microscopist Raymond Royal
Rife, and others too numerous to mention.
This cancer microbe research has been explored in my books The
Cancer Microbe: The Hidden Killer in Cancer, AIDS, and Other
Immune Diseases  and in Four Women Against Cancer: Bacteria,
Cancer, and the Origin of Life -the story of Livingston,
Alexander-Jackson, Diller and Seibert-four outstanding women
scientists who attempted to bring the cancer microbe to the
attention of a disinterested medical establishment. I was
privileged to have met all these remarkable women, who greatly
influenced my own cancer research.
Why is research exploring bacteria in cancer so strongly opposed?
Perhaps it poses a threat to the money interests involved in the
established and orthodox treatment for cancer. Various forms of
cancer treatment include surgery, radiation and chemotherapy.
These therapies might have to be reevaluated if it were proven
that cancer was an infectious disease.
SUGGESTIONS FOR FURTHER INTERNET STUDY
Further information pertaining to cancer microbe research (both
pro and con) can be found by Googling: cancer microbe; bacterial
pleomorphism; cell wall deficient bacteria; "alan cantwell";
"virginia livingston"; "Eleanor Alexander-Jackson"; as well as
other names and key words mentioned in this communication.
For a list of scientific publications pertaining to the
microbiology of cancer, go to the PubMed website hosted by the
National Institute of Health (www.ncbi.nlm.nih.gov) and type in
"Cantwell AR", "Livingston VW", "Alexander-Jackson E", "Diller
IC", "Seibert FB", etc. in the search box.
This short communication is unlikely to convince many health
professionals that bacteria cause cancer. However, after four
decades of studying cancer microbes in cancerous tissue, I am
personally convinced that Dr. Virginia Livingston will one day be
vindicated and recognized as one of the greatest medical geniuses
of the twentieth century.
Ralph W Moss, cancer advocate and author of The Cancer Industry,
notes her passing was "a major loss to the cancer world." In the
Cancer Chronicles #6, 1990, he writes, "Virginia Livingston was a
great person and a great scientist. Sadly, she never received the
recognition she deserved in her lifetime. The true scope of her
achievements will only become known in years to come."
This report honors the centennial of her birth which takes place
on December 28, 2006.
Alexander-Jackson E. A specific type of microorganism isolated
from animal and human cancer: bacteriology of the organism.
Growth. 1954 Mar;18(1):37-51.
Cantwell AR. Variably acid-fast cell wall-deficient bacteria as a
possible cause of dermatologic disease. In, Domingue GJ (Ed). Cell
Wall Deficient Bacteria. Reading: Addison-Wesley Publishing Co;
1982. Pp. 321-360.
Cantwell A. The Cancer Microbe. Los Angeles: Aries Rising Press;
Cantwell A. Four Women Against Cancer. Los Angeles: Aries Rising
Diller IC, Diller WF. Intracellular acid-fast organisms isolated
from malignant tissues. Trans Amer Micr Soc. 1965; 84:138-148.
Greenberg DE, Ding L, Zelazny AM, Stock F, Wong A, Anderson VL,
Miller G, Kleiner DE, Tenorio AR, Brinster L, Dorward DW, Murray
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Viable bacteria present within oral squamous cell carcinoma
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Nuzum JW. The experimental production of metastasizing carcinoma
of the breast of the dog and primary epithelioma in man by
repeated inoculation of a micrococcus isolated from human breast
cancer. Surg Gynecol Obstet. 1925; 11;343-352.
Russell W. An address on a characteristic organism of cancer. Br
Med J. 1890; 2:1356-1360.
Russell W. The parasite of cancer. Lancet. 1899;1:1138-1141.
Sauter C, Kurrer MO. Intracellular bacteria in Hodgkin's disease
and sclerosing mediastinal B-cell lymphoma: sign of a bacterial
etiology? Swiss Med Wkly. 2002 Jun 15;132(23-24):312-5.
Scott MJ. The parasitic origin of carcinoma. Northwest Med.
Seibert FB, Feldmann FM, Davis RL, Richmond IS. Morphological,
biological, and immunological studies on isolates from tumors and
leukemic bloods. Ann N Y Acad Sci. 1970 Oct 30;174(2):690-728.
Shannon BA, Garrett KL, Cohen RJ. Links between Propionibacterium
acnes and prostate cancer. Future Oncol. 2006 Apr;2(2):225-32.
Wuerthele Caspe-Livingston V, Alexander-Jackson E, Anderson JA, et
al. Cultural properties and pathogenicity of certain
microorganisms obtained from various proliferative and neoplastic
diseases. Amer J Med Sci. 1950; 220;628-646.
Wuerthele-Caspe Livingston V, Livingston AM. Demonstration of
Progenitor cryptocides in the blood of patients with collagen and
neoplastic diseases. Trans NY Acad Sci. 1972; 174 (2):636-654.
Young J. Description of an organism obtained from carcinomatous
growths. Edinburgh Med J. 1921; 27:212-221.
Virginia Livingston (1906–1990) was an American physician and
cancer researcher who advocated the unsupported theory that a
specific species of bacteria she named Progenitor cryptocides was
the primary cause of cancer in humans. Her theories about P.
cryptocides have not been duplicated by researchers, and a
clinical trial of her therapy did not show any efficacy in the
treatment of cancer. The American Cancer Society, which did not
support Livingston’s treatment protocol for cancer, categorically
denied her theory of cancer origins.
Virginia Livingston was born Virginia Wuerthele in Meadville,
Pennsylvania in 1906.
Both her father and grandfather were physicians and she also
pursued a degree in medicine. Prior to attending medical school,
Livingston earned three BA degrees in English, history, and
economics from Vassar College. She then attended New York
University, Bellevue Medical College and in 1936, received her
degree in medicine. She was one of four women in her graduating
Shortly after graduation, Livingston became the first female
resident physician at a New York hospital where she was assigned
to treat prostitutes infected with venereal diseases. While there,
Livingston became interested in the study of tuberculosis and
leprosy, and later scleroderma, a disease affecting the tissues
and skin. After studying scleroderma tissues with the darkfield
microscope, she claimed to find an acid-fast organism that
consistently appeared in her slides. Thinking that scleroderma had
some characteristics that were like cancer, Livingston then began
studying malignant tissues and subsequently claimed to find
evidence of acid-fast organisms in every sample. It was this early
research that prompted the young physician to devote her career to
the study of a specific microorganism involved in cancer.
In 1946, Livingston published a paper in which she stated she had
established that a bacterium was a causative agent in
scleroderma. In 1947, she cultured a mycobacteria-like organism
in human cancer and, according to her peer-reviewed paper,
fulfilled Koch's postulates establishing an apparent cause and
effect. In 1949, Livingston was named chief of the
Rutgers-Presbyterian Hospital Laboratory for Proliferative
Diseases in New Jersey where she continued her cancer research.
It was during this time that Livingston formed a lifetime
association with Dr. Eleanor Alexander-Jackson of Cornell
University. Jackson's specialty was the study of mycobacteria and
particularly, the species responsible for tuberculosis. Jackson
had developed specific culture media for growing the microbe and a
technique for observing it known as the "triple stain" because she
felt this microbe wasn't amenable to conventional modes of
culturing and microscopy.
Livingston and Jackson also collaborated on work on the Rous
sarcoma virus (RSV) at Lederle Laboratories. Livingston claimed
that when RSV cultures were passed through special filters
designed to hold back all but the smallest virus particles, she
was able to grow bacteria; this was considered a controversial
claim since bacteria are considerably larger than viruses and are
not supposed to exist in filtered RSV serum. After healthy animals
were exposed to the Rous bacterial filtrates, Livingston and
Jackson claimed that cancerous lesions developed. This finding
led to speculation that such bacteria could be transmitted from
poultry to humans and this became a primary reason Livingston
ordered her cancer patients to not eat poultry while they
underwent her treatment. Scientists have since rejected Livingston
findings, arguing there is no evidence supporting her claim.
In 1956, Livingston published a paper suggesting a causative
bacterium in Wilson's disease. In 1965, she reported isolation
of a variably acid-fast mycobacterium in patients with myocardial
vascular disease. During this time, she also began a small test
trial of anti-bacterial vaccines made from the body fluids of
cancer patients and reported moderate success. Between the
years 1965-1968, Livingston received Fleet Foundation and Kerr
Grants, and continued her investigation into a bacterial cause of
human cancer. She also published a paper describing the presence
of a substance identified as Actinomycin-D which she said could
damage chromosomes and promote cancer.
In 1969, Livingston and her husband Afton Munk Livingston,
established the Livingston-Wheeler Clinic in San Diego,
California, and began formally treating cancer patients. The
therapeutic program included autogenous vaccine made from killed
bacteria derived from body fluids; a low sodium diet consisting of
organic foods, fruits and vegetables high in a substance
Livingston called "abscisic acid"; immune enhancing vaccines
(gamma globulin, BCG) and antibiotics. Livingston prescribed
antibiotics after cross testing them with patients' cultures to
see which had the most antibacterial activity. Livingston also
recommended that patients not consume poultry products based on
her earlier research.
After her husband’s death, she married Owen Webster Wheeler, one
of the first patients she claims to have successfully treated for
head and neck cancer. Shortly after, the clinic was renamed the
Livingston-Wheeler clinic. In 1970, Livingston officially named
her cancer organism Progenitor cryptocides, and presented her
findings to the New York Academy of Sciences. According to her
biography, Progenitor was a pseudonym meaning "ancestral" and the
name was chosen because Livingston believed the microbe existed as
early as the Precambrian era, and it was an endegenous component
of life itself. The name "cryptocides" was a Greek and Latin word
which meant "hidden killer". The microbe was classified under the
order Actinomycetales. Livingston described Progenitor as an
intermittently acid-fast mycobacterium that displayed highly
variable growth cycles. According to Livingston the microbe was
pleomorphic, and had cell wall-deficient and filter-passing forms
resembling viruses, with the ability to adopt a variety of shapes
including spindles, rods and cocci.
In 1974, Livingston published a paper which described her
isolation of human chorionic gonadotropin (hCG) from cancer
bacteria. She then advanced one of her central hypotheses.
Livingston theorized that hCG is both a component of human cancer,
but also innately involved in embryonic growth and fetal
survival. She wrote that hCG is saturated in the placenta, and
blocks the mothers’ antibodies from attacking the fetus, partly
made of foreign DNA (and not recognized by host immunity). By the
same token, hCG performs a similar function in cancer, conferring
protection to malignant tissues. Livingston believed that after
Progenitor hybridizes with cancer cells, it imparts an ability for
them to produce hCG in a manner similar to that of the developing
fetus. Based on this duality of function, Livingston called hCG
“the hormone of life and the hormone of death”. She also
stipulated that vaccines which target hCG-producing bacteria could
also halt the progression of cancer. And she claimed that absicins
could also neutralize hCG.
Though some bacteria have been associated with cancer (for
instance H. pylori has been associated with stomach cancer)
Livingston's postulated relationship between cancer and P.
cryptocides was never proven in several follow up studies
conducted by independent investigators. Researchers confirmed that
bacteria provided by Livingston produced hCG, but several other
studies demonstrated that numerous bacteria in both cancer
patients and healthy individuals also produced the substance.
Occurring before the existence of techniques to analyze DNA,
Livingston and other investigators' ability to differentiate
bacteria based on morphology and chemical characteristics was
limited. However, even given technological limitations at the
time, Livingston's classification methods were described as full
of "remarkable errors", attributing characteristics to
Actinomycetales (the order Livingston believed P. cryptocides
belonged to) shared by no other members of the order. Some
evidence supports P. cryptocides is the result of a mistaken
identification of a Staphylococcus strain of bacteria and later
studies of the samples provided by Livingston proved to be
Staphylococcus epidermidis and Streptococcus faecalis.
The American Cancer Society (ACS) did not support Livingston’s
treatment protocol for cancer, and has categorically denied her
theory of the cancer bacterium P. cryptocides the primary cause of
human cancer. The ACS also challenged the efficacy of Livingston’s
autogenous vaccine and concluded in its report that there was no
corroboration of either P. cryptocides or the efficacy of her
autologous vaccine. Since Livingston hadn’t stocked earlier
cultures of her alleged microbe, it is not possible to decipher
precisely what those cultures contained.
A case-control study using self-selected, matched but not
randomized groups with late stage cancer compared survival and
quality of life between cancer patients receiving conventional
treatment and those undergoing the Livingston-Wheeler therapy. The
results were reported in The New England Journal of Medicine in
1991, and found no differences in survival among patients whether
treated conventionally, or via Livingston's treatment. The NEJM
report also stated that when comparing the two groups, the
"quality of life were consistently better among conventionally
treated patients from enrollment on". Based on this trial, the ACS
deemed Livingston's cancer therapy without efficacy, and
considered it an "unproven therapy".
While both groups of patients in the trial deteriorated at equal
rates---all in effect dying of their disease---patients in the
Livingston-treated group were reported to have had a "poorer
quality of life" at the start of the trial. The study's lead
investigator, Barrie Cassileth, acknowledged that "the University
of Pennsylvania patients had a significantly better quality of
life at all times, including enrollment" and that, quality of life
"was different at base line", with Livingston's patients rated
worse. Patients in both treatment arms also received
conventional therapies in addition to Livingston's therapy.
Livingston's patients also received BCG during the trial---an
FDA-approved cancer adjuvant which has been found effective for
several cancers, including those of the bladder and colon.
At the study's conclusion, Barrie Cassileth commented:
"This study...involved only patients with diagnoses and stages of
disease for which there is no effective conventional treatment.
Therefore, the results cannot be generalized to patients with less
advanced stages of disease or to other treatment regimens."
Cassileth also said, her study group "hypothesized that survival
time would not differ between the two groups on the basis of the
assumption that the unproved remedy would be no more effective
with end-stage disease than conventional care, itself largely
Shortly after speaking before an Office of Technology Assessment
(OTA) hearing on alternative cancer therapies and attending her
60th reunion at Vassar College in 1990, Livingston accompanied her
daughter Julie Anne Wagner on a European trip. She developed chest
pains while visiting the Greek islands and then succumbed to heart
failure in Athens on June 30, before being transported to a Paris
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March/April 1990, Pages 103–108
Unproven Methods of Cancer Management :
CA Cancer J Clin. 1990 Mar-Apr;40(2):103-8.
Livingston-Wheeler's cancer treatment is based on the belief that
cancer is caused by a bacterium she has named Progenitor
cryptocides. Careful research using modern techniques, however,
has shown that there is no such organism and that
Livingston-Wheeler has apparently mistaken several different types
of bacteria, both rare and common, for a unique microbe. In spite
of diligent research to isolate a cancer-causing microorganism,
none has been found. Similarly, Livingston-Wheeler's autologous
vaccine cannot be considered an effective treatment for cancer.
While many oncologists have expressed the hope that someday a
vaccine will be developed against cancer, the cause(s) of cancer
must be determined before research can be directed toward
developing a vaccine. The rationale for other facets of the
Livingston-Wheeler cancer therapy is similarly faulty. No evidence
supports her contention that cancer results from a defective
immune system, that a whole-foods diet restores immune system
deficiencies, that abscisic acid slows tumor growth, or that
cancer is transmitted to humans by chickens.
Ann. N.Y. Acad. Sci., Volume 174, Unusual Isolates from
Clinical Material, Pages 675–689 ( Oct 1970 )
TOXIC FRACTIONS OBTAINED FROM TUMOR
ISOLATES AND RELATED CLINICAL IMPLICATIONS*
Annals of the New York Academy of Sciences. 174 (2):
A specific type of organism
cultivated from malignancy: bacteriology and proposed
Livingston, VW; Alexander-Jackson
Transactions of the New York Academy of Sciences. 36 (6):
immunological, and biochemical properties of Progenitor
Livingston, VW; Livingston, AM
Guinea pigs became skin-positive to test doses of PPD
(purified protein derivative-Seibert) and to other related PPD
preparations in various degrees following immunization with
phenolized cultures of Progenitor cryptocides, an intermittently
acid-fast pleomorphic filterable Actinomycete isolated from human
cancer patients. This reactivity, indicative of the relationship
to M. tuberculosis as well as to several other related microbes,
may account for the effective treatment of some types of human
cancer with BCG (Bacillus Calmette Guérin). Another property of P.
cryptocides is the production in vitro of a parahormone
immunologically and biologically related to human chorionic
gonadotropin. Since the cancer patient often exhibits various
types of hormonal imbalance, the microbic exogenous production of
this hormone may explain the neoplastic parahormone syndrome in
man. There are some biological differences in experimental animals
between the microbic and human chorionic gonadotropin, but the in
vitro and radioimmunological tests are identical. The microbic
hormone appears to play an important role in human cancer, since
it not only is present in tumors, body tissues, and fluids but may
be excreted in large amounts in the urine. The amount excreted is
variable, depending upon the rate of microbic production of the
hormone in vivo and the resistance of the host as expressed
through immunological and metabolic degradation systems.
During the last hundred years, a surprisingly large number of
doctors have found ways to cure or alleviate cancer. These
ways are not the Big Three – surgery, radiation, and chemo – that
are the ones almost exclusively recommended by the profession.
Let’s begin with Virginia Livingston (1906-1990), a graduate of
Bellevue Medical School, daughter of Herman Wuerthele, MD
(1885-1967). In her first book, Cancer: A New Breakthrough (1972),
she claimed a success rate of 82%. Here are four cases:
D.K. – Age 71, operated on for carcinoma of prostate, followed by
removal of testicle, 1966. He had multiple spinal metastases and
arthritis of many joints. He was barely able to move around. He
was placed on autogenous vaccine and mandelamine, 1 gram four
times a day with dietary and vitamin adjuvants. Previous to his
prostatic surgery he had a bowel resection for cancer of the
colon. At the present time the spinal metastases have healed, he
says he has no evidence of arthritis, is in perfect health and
J.M. — Age thirty-five, had a left radical mastectomy March 3,
1965, when four months pregnant. Pathological diagnosis was
infiltrating arcinoma, scirrhus and medullary types. After
delivery of a normal child she had a hysterectomy May 28, 1965,
and was placed on estrogen therapy from August 24, 1966, through
January 9, 1967. Autogenous vaccine was made which she took for a
year and intermittently since. This type of tumor is universally
fatal. Her physician says she is in good health at the present
time (1972) with no signs of recurrence.
F.B. — Male age twenty-seven from Utah, who was operated on for
severe headaches after a number of convulsive seizures. The
pathological diagnosis was astrocytoma, grade III to IV,
infiltrating the surrounding area. He received anti-convulsants,
radiotherapy and antibiotics. In 1966 when he was doing very badly
and appeared to be terminal, he was placed on autogenous vaccines
and mandelamine, one gram four times daily, plus vitamins and
dietary supplements. He remained on this regimen for two years.
The vaccine was discontinued in October 1970. His physician said
there is no evidence of any tumor.
A Longshoreman, age 46, operated 1967, for a mass on the right
side of his neck. Pathological diagnosis was malignant lymphoma,
reticulum-cell type with invasion of all glands. These were not
resectable because they extended under the sternocleidomastoid
muscle. He received X-ray, 4500 R, in eighteen treatments. Since
then he has had no other treatment except autogenous vaccine
continuously with erythromycin 250 mgm twice a day. He says he is
completely well and works full time on the docks.
Leprosy the Clue
After World War II, Virginia worked in a New York hospital and saw
many cases of TB (tuberculosis) and leprosy. Note: every
physician’s experience is unique. It is incorrect to think that
all doctors possess the same knowledge; much depends on who
happens to walk into their office one fine day.
One fine day into Virginia’s office (she was a school doctor)
walked the school nurse, complaining of ulcers on the fingers, a
perforation in the septum (the piece of cartilage that separates
the two nostrils), and hardening of the skin. This was in 1947.
Her own doctor had given her a diagnosis of scleroderma.
Virginia associated the symptoms with leprosy as the patient
reported that she could not feel hot or cold on the affected skin.
Virginia Livingston decided to do some lab work on this case. She
took smears from the woman’s nose and the ulcers on her hands and
stained them with the stain used for identifying both leprosy and
TB, namely a “Ziehl-Neelsen” stain.
Peering into the microscope, Virginia saw the same type of
microorganisms one sees in leprosy. She treated this patient with
the medication used for lepers, and the skin healed. Later,
Virginia gave the same medication to other scleroderma patients
and it worked!
Whatever she saw in the microscope that day became central to her
later theory that cancer is explainable by bacteria. That has not
been widely accepted. But she made a separate discovery that did
later become standard in science. Namely, she found that bacteria
can and do secrete a hormone, human chorionic gonadatropin, hCG,
which is essential for human life. Hooray!
In a Nutshell, Livingston’s Theory
Virginia believed that cancer is not a foreign visitor. It is part
of our body from birth and it is never going to go away. Cancer is
characterized by mitosis, the dividing and replication of cells.
Cell division itself is not to be despised; it is the basis of our
initial growth in childhood, and occurs as part of the repair work
that steadily goes on in the body. When a piece of skin gets
scraped off, you just wait for it to regrow. We need cell
If cell division gets out of control, however, it may make tumors.
Tumors are bunches of new cells that don’t associate in the normal
manner with surrounding cells and have no purpose. A cancer doctor
has the title “oncologist” from the Greek word onco for mound.
Virginia never became a “moundologist.”
She surmised that a tumor happened because the person’s immune
system was not functioning as it normally does. As for the cancer
microbe that she believed to be ever-present in our body, she gave
it the name Progenitor cryptocides (crypto=stealth; cide=to kill).
The Livingston Program for Treating Cancer
Virginia Livingston does not claim to have invented the bacterial
theory of cancer. Others such as William Russell and Royal Rife,
she notes, got there first.
Now have a look at what she prescribes: she tells the cancer
patient to get his Progenitor cryptocides back under control. That
is something that, in a healthy person, is taken care of by the
When your immune system sees the cryptocides microbes going where
they shouldn’t go, she says, it treats them as invaders and acts
to protect you. The immune system is ever-alert for the
non-normal, and can do what must be done.
One cancer patient, a physician named Owen Wheeler, was cured by
Virginia, and subsequently married her. They established the
Livingston-Wheeler Clinic in San Diego, and helped thousands of
But what if your immune system is not working well and can’t call
up the right response? Then a tumor may form. Stuff may also
travel around your body and metastasize.
What should the doctor do? She will try to get your immune system
Livingston’s treatment program has two prongs:
Use nutrition to build up the immune system. She advises fresh
fruits, vegetables, and nuts (nothing out of a can). No meat or
dairy until you are recovered. Lots of Vitamins A and C, and
Vaccinate the patient with the antigen he needs. Material for that
vaccination comes from the patient himself; his urine is used to
culture the bacteria which are then made into an autogenous
vaccine. In some cases she also gives antibiotics. She often gives
a blood transfusion, from a family member.
“Getting” Virginia Livingston
Virginia was still working at her clinic at age 83 when the
government closed it down. The feds and most states do that to any
doctor who dares defy the rule to use only the Big Three cancer
cures. (surgery, chemo, radiation).
A few months after that, she expired.
Dr Livingston constantly made her patient’s progress available for
inspection by the medical authorities. She also arranged for a
random survey of the records, going so far as to hire an outsider
to choose 62 cases under a meticulous set of guidelines. Yet when
she published the survey no one was wiling to read it.
In the back of her book you will find a section with the pitiful
heading “Ten Cases That I Wish Someone Would Investigate.”
In 2001, Saul Green wrote a bad evaluation of Livingston for Sloan
Kettering, making nary a mention of cures she wrought for 20
years! He did, however, importantly remind us that it is a felony
in California to treat a cancer patient with unapproved methods.
Cantwell and Pleomorphism
One physician who came to Livingston’s aid was Los Angeles
dermatologist Alan Cantwell, MD.
He had already published his clinical finding of a cancer microbe,
in 1968, before he got to know Livingston. Cantwell spends much
time at an ordinary microscope using an “oil immersion lens” that
allows him to see what most pathologists claim they don’t see. He
credits a Spanish microbiologist for giving him the clue to
cancer’s similarity to tuberculosis:
“[Conrado] Xalabarder totally transformed my concept about how
tuberculosis-causing mycobacteria reproduce and grow and
drastically change their appearance.”
Ah, changing appearance – and behavior – and size, and – who
knows? Maybe their species identity – is the name of the game for
the bacteria we are concerned with here.
The pioneering taxonomist, Carolus Lineaus, born 1707 in Sweden,
grouped animals together in phyla based on shared characteristics,
for example, he put Homo sapiens into the phylum chordata, as we
share the characteristic of having a backbone. (Sigh. If only we
had backbone, and not just a backbone!)
When a bacterium is not encased in the normal way by a cell wall
(that is, when it’s CWD – cell-wall deficient), it has potential
to sneak around and get up to no good. Harken to this:
“There is increasing evidence that CWDB and CWD fungi are often
associated with endocarditis, septicemia, meningitis, pneumonia,
and infections of bones and joints. When prompt diagnosis is
critical, it is helpful to include examination for CWD
microorganisms as part of the first laboratory study.”
That’s from an article that Lida Mattman, and her co-author Mehnga
S. Judge, contributed to Domingue’s Cell Wall-Deficient Bacteria.
(1982: 440). Amazingly, it did not spark eager research as to its
rather glaring potential for meningitis cases.
Patent #4, 692, 412 (expiry date 2006) was awarded to V.
Livingston, and Afton Livingston, and Eleanor Alexander-Jackson
for the making of an autogenous vaccine from the blood or urine of
cancer patients. Their application for the patent claimed:
“All cancerous bloods examined have revealed the cryptocides
organisms. [It] is apparently ubiquitous in nature, existing in a
reservoir in soil and water, and is found in all classes of
animals…. It can exist as a latent infection in host tissue
without causing apparent ill effects. However, when the
immunologic barriers are lowered it can invade the host in
prodigious numbers and involve any or all of the host tissues,
causing various kinds and degrees of pathologic change.” [Note:
Award of patent does not mean claim proven.]
This Patent’s Method for Making Autogenous Vaccine:
“Obtain a midstream clean-catch specimen of urine in a sterilized
screw-top glass container. Make up DiFco’s brain-heart infusion
agar: 37 grams of the agar base are added to a liter of distilled
water heated to melt and mix, and distributed into flasks or
bottles of 95 ml amounts, and autoclaved. Five percent (5%) human
blood … is added when the melted agar has cooled down to 45-50
degrees C., and the mixture is poured into sterile Petri dishes.
Streak the surface of the blood agar plate with a sterile swab
dipped in the urine. Incubate plate to 37 degrees C. and examine
after 24 hours.
“If growth has appeared, note types of colonies, make duplicate
smears, and stain one by Gram’s stain and the other by
Alexander-Jackson’s modified Ziehl-Neelsen technique: flood slide
with Kinyoun’s carbolfuchsin for 3 to 5 minutes in the cold, wash,
decolorize briefly with 70% alcohol containing 1 to 3% HCl as
these organisms decolorize more readily than M. tuberculosis,
counter stain by flooding slide with Loeffler’s methylene blue and
add 6 to 8 drops of normal (4%) sodium hydroxide. Tilt slide to
mix, and wash after 30 seconds.”...
Livingston-Wheeler Therapy is an alternative approach to cancer
treatment that gained popularity from the 1970s to early 1990. It
used several treatments to supposedly stimulate the immune system,
including a strict vegetarian diet, BCG vaccine, coffee enemas,
autogenous vaccine, vitamins, antibiotics, antioxidants,
nutritional counseling, and support groups/counseling. The regimen
was based on the theory that cancer was caused by the bacterium
Progenitor cryptocides, which developer Virginia
Livingston-Wheeler reported to have isolated in a wide variety of
cancer tissues (3) (4). A weakened immune system would then allow
the bacterium to grow, and consequently the therapy’s focus was
immune-stimulation. Although a number of viruses and bacteria have
been associated with various cancers (8), a link between the
bacteria named by Livingston-Wheeler and cancer was never
confirmed by independent researchers.
A self-selected, matched-cohort, prospective comparison of
patients at Livingston-Wheeler Clinic and a conventional cancer
center found no difference in survival times between groups, but
did find consistently lower quality of life in the
Livingston-Wheeler cohort (6).
Metabolic diets may result in nutrient deficiencies (5). Repeated
use of coffee enemas has been linked to several deaths from
serious infection and electrolyte imbalance (2). In a 1990
position paper, the American Cancer Society urged cancer patients
not to use these treatments (1).
Mechanism of Action
Although a number of viruses, bacteria, and parasites have been
associated with various cancers (8), a link between the bacteria
named by Livingston-Wheeler and cancer was never confirmed by
independent researchers. Rather, independent analyses of cultures
provided by Livingston-Wheeler identified these bacteria as Staph
epidermis, Strep faecalis, Staph faecalis, and other unrelated
In a 1990 position paper, the American Cancer Society urged cancer
patients not to use this treatment, as no evidence supports its
efficacy (1). In addition, although the Livingston-Wheeler diet
was similar to recommendations made by the American Cancer
Society, its nutrient deficits, especially for calcium, iron,
vitamins D and B12, and protein, would be unsuitable for some
Common (metabolic diet): Nutrient deficiencies (calcium, vit B12,
protein), anemia, and malabsorption may result from metabolic
Reported (autogenous vaccine): Malaise, aching, slight fever, and
tenderness at injection site (1).
Case Report (Coffee enemas): Death attributable to fluid and
electrolyte imbalance causing pleural and pericardial effusions
after use of coffee enemas, 4 per day for 8 weeks (2).
The Woman Who Cured Cancer: The Story of
Cancer Pioneer Virginia Livingston-Wheeler, M.D., and the
Discovery of the Cancer-Causing Microbe
Edmond G Addeo
This story is now more relevant than ever as the latest science is
now validating the protocols of Dr. Livingston-Wheeler who will
one day be placed in the same class as Pasteur, Curie, Salk/Sabin
and their discoveries.
The Conquest of Cancer
by Dr. Virginia Livingston-Wheeler (Author), Edmond G. Addeo
This is the original book published in 1985 on the pioneering work
of Dr. Virginia Livingston-Wheeler. It is being republished to
make it widely available to the general public.
Quotes from the original book: "Dr. Virginia Livngston-Wheeler is
one of the great, unsung scientists of present day medicine. When
her discovery of the 'cancer microbe' becomes... accepted, she
will undoubtedly be known as the Pastuer of this century." Alan R.
Cantwell, Jr. M.D.
"Dr. Virginia has long been pioneering in new concepts of cancer
cause and treatment... the many successess she has had in treating
advanced cancer patients demonstrate what can be accomplished when
a physician in cooperation with understanding patients tries to
expand the limits of cancer treatment." Ray G. Crispen, Ph.D
"This important book could very well save your life or the life of
someone you love. Dr. Virginia Livingston-Wheeler has succeeded in
devising a scientifically rational program for the prevention and
treatment of cancer." Richard A. Kunin, M.D.
Method of preparing an autogenous vaccine
Inventor(s): LIVINGSTON VIRGINIA, et al.
A method of preparing an autogenous vaccine for use in improving
the immunocompetence of animals affected with neoplastic disease
characterized by the production of choronic gonadotropin by the
microorganism Progenitor cryptocides, ATCC 31, 874.
DESCRIPTION OF THE INVENTION
This invention relates to chemical substances comprising a product
of the growth of a microorganism of Order II, Actinomycetales
Buchanan, as identified in Bergey's Manual of Determinative
Bacteriology, Sixth Edition, 1948, p. 111, to method of preparing
same and isolation thereof and to the chemotherapeutic treatment
of animals and humans therewith. Published works have identified
additional species and resulted in a classification of the
organism under the order Actinomycetales. This order has been
reclassified as follows:
Cryptocides sclerodermatis (sclerobacillus)
stains isolated from lupus erytheuratosis, rheumatoid, arthritis,
periarthritis, nodosum, sarcoidoses (that is, from collogen
diseases other than cancer and other such diseases specified
varieties: hominis, rodentii, avii, etc. A culture of Progenitor
cryptocides was deposited in the American Type Culture Collection,
Accession Number 31,874.
All of the species noted above have been observed to be
interchangeable within the scope of this invention.
P. Cryptocides has been assayed and assigned the tentative
formula, C30 H38 N2 O3 (certain products of Cryptocides were
crystals extracted from cultures of urine from terminal cancer
patients). Extracts also have been crystallized from the blood and
urine of cancer patients and the crystals result from the presence
of the organism.
The microorganism involved (Cryptocides) has been identified as a
highly pleomorphic intermittently acid-fact micro-organism, with
both a virus-like and a PPLO or L transitional phase. This
organism is a great simulator, whose various forms may resemble
micrococci, diphtheroids, bacilli, fungi, viruses, and host-cell
inclusions. Cryptocides has the ability to change its form and may
vary its appearance from that of a fungus to that of a cluster of
virus-size pleuro-pneumonialike organisms (PPLO or Mycoplasma).
Collagenophillic mycobacterium-like, which include the cancer
organism, are able to change their forms. Cryptocides has
filterable or extremely small forms (submicroscopic) similar to
viruses, and rather large mycelia. There were some variations as
to size and some differences in the kind of media or material in
which it will grow. Certain strains of it ferment one kind of
sugar, some others, and some can live with little (or no) oxygen,
whereas some require more. It can be identified as a single agent.
The microorganism undergoes many changes in morphology and some of
those forms might be zoogleal or "L" Forms. Zoogleals are
intermediate forms of microorganisms which ordinarily have cell
walls, but, in which, under certain circumstances the cell walls
are absent. Some of these forms can be passed through very fine
filters that hold back the usual bacteria and allow only very
small particles such as viruses and small L forms to pass through.
Such filter-passing bodies can regrow to become bacilli (bacterial
cultures). The microoganisms involved have many forms but they
always grow up to be the same thing no matter how often they are
cultured. Cryptocides is not a virus but is a pleomorphic
Cryptocides is acid fast, that is, it retains the Ziel-Neelsen
stain in the presence of acid. Cryptocides is related to the
tuberculosis family of microbes. It is filterable through filters
designed to hold back bacteria. It is sensitive to tetracycline,
kanamycin, ampicillin and furacin, but occasionally resistant to
pencillin, sulfa drugs and mycostatin. As for the pleomorphism,
cryptocides exists as virus-sized bodies of 20 to 70 microns, as
elementary bodies of 0.2 micron, and in coccoidal forms of 0.5
microns or larger. The latter are usually gram-positive and
resemble common micrococci but are distinguishable by variation in
size and the sprouting of filaments or spicules. The organism may
also appear in amorphous mycoplasmalike forms, as rods or
filaments of varying lengths, and in older cultures, as spores and
Cultures made from animal tumors and fluid have great similarities
with cultures derived from many types of fresh uncontaminated
human tumors, from blood and other body fluids of patients who
have advanced cancer.
The L-forms are bacterial forms without cell walls. They resemble
pleuro-pneumonilike organisms (PPLO), also known as mycoplasma.
However, the mycoplasma appear to reproduce continously under some
conditions in the same stage, with the absence of cell walls,
while other organisms have a tendency to revert more quickly to
the more stable bacillary or coccal forms of origin. The L-forms
are the link between bacteria and the viruslike minute bodies that
are a stage in the life cycle of certain microorganism. Many
viruses may actually be L-forms of microbes which, under certain
conditions may be induced to return to their original forms.
Previously the appearance of the both adult and L-forms led to the
erroneous conclusion that there was a mixture of microorganisms, a
contamination of pure strains with other nonrelevant
microorganisms, but this was shown to be an erroneous
misconception. Some true contaminants are readily recognized by
their growth pattern but the Cryptocides is a great simulator of
other organisms. It requires infinite patience to observe its
growth pattern and to recognize its transition from one form into
The microorganism involved requires definitive bacteriologic media
for its primary isolation, differential staining techniques for
its identification, high power microscopic resolution, and the
electron microscope to reveal its most minute forms. Specific
cultures can be obtained on solid media used for the isolation of
the tubercle bacillus. This mycobacterium-like organism is
believed to be a primary etiologic agent in proliferative and
degenerative diseases such as cancer and perhaps many other
socalled autoimmune diseases. Cryptocides is believed to be the
causative or infectious agent (microbial) of cancer, in all of its
forms, Cryptocides would therefore be called an antigen. More
specifically, the filterable forms of P. Cryptocides which are of
virus size are the causative agents in human and animal cancers.
Peyton Rous did not call his tumor filtrates (from chickens)
viruses but instead "tumor agents." His material could be dried
and held on a shelf at room temperature for months and then, mixed
with saline, it could be reactivated to initiate fresh tumors. A
true virus has been defined as a submicroscopic infectious unit
that lives only in the presence of living cells and cannot exist
even momentarily outside of them. Many have tried to find a virus
implicated in some form of human cancer, but none has been found.
Applicants have grown the Rous tumor agent in sterilized beef
broth that contained suitable nutrients for bacterial growth, and
traced its growth pattern through all of the bacteriological
stages. Applicants knew that the infectious agent passed through
filters that permitted only the passage of so-called viruses.
Applicants filtered the cultures, not the extracts of the tumors,
through bacteria restraining filters and studied these with a
electron microscope. Applicants kept the cultures in which there
did not appear to be any visible form of life, incubated them at
37 DEG C. and from these seemingly clear broths with the agent in
them, there arose the bacterial and fungal stages of the
cryptocides. Applicants ruled out contamination that might account
for the bacterial growth on incubation by repeating the experiment
dozens of times. It was a tedious process but proved that this
so-called virus could and did convert to a bacterium that had not
only submicroscopic forms but also bacillary, coccal or round
forms, and that could also develop funguslike stages and spores.
(On studying the growth of the tubercle and lepra bacilli these
stages were entirely comparable with the Cryptocides.)
When applicants examined the cultures obtained from human cancers,
there was no discernible difference in the growth pattern. The
growth pattern of the chicken cancer isolates and that of man were
the same. They grew in the same kind of broth in the same way and
they appeared the same in chicken and human tissues. They had the
same staining properties with the Ziehl-Neelsen dye. Applicants
did sheep immunization studies in which they found significant
cross-agglutination between the Rous sarcoma, fowl leukosis and
various strains of human cancers. When applicants injected the
isolated cultures into mice, the characteristic disease and
lesions developed. The Rous isolates had to be readapted to
chicken tissues by passage on the allantoic membrane of fertile
eggs and then replanted into young chicks. Applicants also carried
on immunization of rabbits with the leuokosis agent and used the
antiserum to cure chickens dying of fowl leukosis. In every way
the Rous agent appears to be a prototype for human cancer.
The above studies led to the cultivation of the same kind of
microorganisms from other animal tumors. These invariably grew and
appeared similar to the Rous and human strains. Sometimes there
were differences in size or different sugar or oxygen requirements
for cultivation, but essentially they were the same basic type or
The cancer organisms (cryptocides) appear to resemble mycoplasma,
organisms that exist without cell walls, expecially since the
cytosinguanine ratio of their mucleic acid, DNA, is similar to
that of certain mycoplasma. However, the usual mycoplasma tend to
remain in their state of existence without cell wals but the
Cryptocides may pass rapidly through the stage without walls to
the form of true bacteria. Perhaps all mycoplasma could be induced
to become bacteria but this is still a disputed point.
Dr. Robert Huebner, head of the National Institute of Allergy and
Infectious Diseases, Bethesda, Md., has stated that cancer is a
viral infection. Of the various agents suspected, he stated that
the C-particle is the most likely agent. It has been called by
this name because the round bodies found in cancerous tissues
often appear in a C shape. However, the comparison of the
C-particles in mouse leukemia with the filted Cryptocides isolates
examined under the electron microscope show them to be similar in
size and shape. In preparations from cultures the round forms are
often seen to split and assume the C form. It would seem that this
splitting into a C is characteristic, but not necessarily a method
of identification. All the other methods are necessary as well.
The cancerous growth itself is not the entire disease. The small
coccuslike granules which can be seen dividing in cancer cells
represent the intracellular parasite that is the causative agent.
The parasite within the cancer transforms the normal cell into a
sick cell that cannot mature by differentiation. It is the
filtered material from tumors and other cancerous growths, as well
as the cells themselves, which transmit cancer from one species to
another species. Or, in other words, the cancerous agent can cross
species barriers and infect other species.
P. Cryptocides not only causes cancer but a number of other
ailments that effect man. The infectious nature of arthritis, of
some kinds of heart, liver and kidney impairment, and most
recently of diabetes is known. The patterns of these diseases
point to their latent infectious nature. Cryptocides is an
infectious agent. But, the tumors are only a part of the resultant
disease. In addition to tumors, there are cheesy lesions or areas
resembling tuberculosis, which can involve any one of the
essential organs such as the liver, kidney heart or lung. These
organs might show changes in the connective tissue, called
collagen, which can lead to degeneration as seen in the chronic
human degenerative diseases. The organism may assume a latent form
and be inactive as long as the body's defense mechanisms are
adequate, but when they are not, disease results. The exact kind
of disease depends on the age of the host and its state of
resistance, as well as the strain of the organism.
All cancerous bloods examined have revealed the cryptocides
organisms. Applicants have made a film of untreated blood from a
terminal cancer patient in which the parasites are seen in
Brownian motion in the red cells. The parasites stay inside the
cells of patients who are holding their own against the disease,
but in advanced cases, the numerous cells rupture, releasing the
The microorganism is apparently ubiquitous in nature, existing in
a reservoir in soil and water, and is found in all classes of
animals, including man. It can exist as a latent infection in host
tissue without causing apparent ill effects. However, when the
immunologic barriers are lowered it can invade the host in
prodigious numbers and involve any or all of the host tissues,
causing various kinds and degrees of pathologic change
equilibrated between the ferocity and numbers of the invader and
the ability of the host to resist them. Not only is the organism
pleomorphic but the pathologic changes induced in experimental
animals show varying degrees of disease ranging from the lethal
through the semi-immune, neoplastic and degenerative stages.
Hyper-immune and degenerative stages may be relatively quiescent
but can become slowly and progressively fatal.
Man and/or animal can be a latent carrier of the Cryptocides. Many
of applicants' experimental animals that have surived cancer have
developed interstitial collagen disease as a result of their
inoculations with applicants' bacterial isolates of Cryptocides,
and also developed heart lesions. When baby mice born of infected
mothers died, the autopsy showed destruction of heart muscle.
These lesions contained the acid-fast organisms (Cryptocides) in
the heart muscle. Also, a number of research people in England
have reported strange microbic bodies previously unrecognized in
the hearts of people who died of coronary disease. One of the
applicants had cancer of the forehead treated successfully with
radium fifteen years previously, but was a latent carrier of the
Cryptocides. That applicant was treated after that period of time
with an autogenous vaccine, has had a new vaccine prepared every
year, and has continued treatment.
"Mycobacterial Forms in Myocardial Vascular Disease", Virginia
Wuerthele-Caspe Livingston and Eleanor Alexander-Jackson, (1965)
proposes the theory that there are microbic bodies in the lesions
of heart diseases and that they are especially numerous in the
areas where the blood vessels have ruptured. Until recently the
theory has been the coronary blood vessels of the heart are
narrowed due to arteriosclerosis, and that fatty deposit in the
wall of the vessels, and overweight are the determining factors in
this type of heart disease. Now the medical researchers are
becoming aware of the fact that the blood bessels themselves are
often not involved so much as the supporting tissues and muscles
of the heart so that the heart vessels rupture due to extrinsic
factors outside the vessel rather than from intrinsic disease.
This is particularily true of patients with collagen diseases such
as scleroderma and lupus erythematosis. Vascular and myocardial
pathology is related to chronic low-grade infection by the
mycobacterium-like organisms (Cryptocides).
Degenerative changes occur in coronary heart disease in the
presence of the invasive mycobacterial parasite cryptocides.
Postmorten heart sections of 6 patients with coronary and aortic
disease were stained by the Fite modification of the Ziehl-Neelsen
technique (for demonstrating Lepra bacilli in sections) using
Kinyoun's carbon-fuchsin, and compared with sections of the same
involved areas stained with conventional H and E. Eight
predominant types of lesions were observed in the myocardium
1. PERIVASCULAR CHANGES AROUND THE SMALL CORONARY VESSELS. In the
loose connective tissues numerous small acid-fast bodies can be
2. CELLULAR INFILTRATION. This is frequently seen not only around
the vessels but between the muscle fibers as well. These cells
consists almost entirely of mononuclear types, predominantly
lymphocytes, while large mononuclear phagocytes laden with
organisms plasma and other mononuclear cells are present in
relatively large number.
3. FIBROBLASTIC INFILTRATION. The presence of these organisms
appears to stimulate the formation of fibroblasts. In some areas,
the muscle fibers and interstitial tissues appear to be replaced
4. INFARCTION. Where there has been an infarct, there may be a
softened central area with numerous small acid-fast cocci and
coccobacilli present in the collagenous hemorrhagic softened area.
5. NECROSIS. Necrotic changes may involve the blood vessels.
Striking degenerative changes of the vessel walls are observed as
illustrated not only by the sections of coronary vessels but also
by the sections of in involved aorta. Proliferative changes may
involve the endothelium, with invasion of the endothelial cells,
and are accompanied by thickening and narrowing of the wall.
Hairlike filaments of the organisms were seen protruding into the
lumen. These changes are also present in the vasa vasorum of the
6. THROMBOSIS AND RECANALIZATION. Some areas of recanalization
were observed in heart, liver, and spleen.
7. CHANGES IN THE ELASTIC LAYER OF THE AORTA. The elastic fibrils
have lost their identity and have become collagenized with loss of
structure. As scar tissue forms, cholesterol-like plaques occur.
It seems possible that deposity may be derived in part from the
fatty envelopes of these organisms. In other tissue where masses
of the organisms have proliferated, polyhedral crystals resembling
cholesterol have been observed.
8. CHANGES IN THE HEART MUSCLE. Individual nuclei of the heart
muscle are frequently parasitized, and replaced by small acid-fast
globoidal bodies. The muscle fibers themselves appear in a state
of gradual digestion and disintegration by both minute and larger
All of the above can be treated and detected by this invention
Neoplastic changes have been shown by Diller and Diller
(Intracellular acid-fast organisms isolated from malignant
tissues, Trans. Amer. Micr. Soc., 84:138-148, 1965), to arise in
tissue culture as the result of exposure to this specific invading
The Journal of the American Medical Association, July 28, 1969
Vol. 209, No. 4, contains a summary of the work of K. A. Bisset,
New Scientist, June 12, 1969, who speculates that many diseases
like leukemia and arthritis could be caused by Mycoplasma or by
forms of this elusive bacteria and wrote that the fact that
mycoplasm can break down into viruslike particles, easily
identifiable on electron-microscope examination and similar to
those found in blood of leukemia patients, leads to a strong
suspicion that Mycoplasma may be a culprit in the development of
certain malignant processes.
Dr. Florence B. Seibert, Veterans Administration Research
Laboratory at Bay Pines, Fla., has reported immunologic studies
with the organisms. Labeled antiglobulin, which was specific for
an isolate from a human breast, adenocarcinoma induced specific
fluorescence in the white blood cells of patients with leukemia
and myeloma, demonstrating an immunologic relationship.
Koch's law is the foolproof method of proving the cause of a
disease. It is as follows:
1. The microorganism must be present in every case of the disease.
2. It must be possible to cultivate the microorganism outside the
host in some artifical media.
3. The inoculation of this culture must produce the disease in a
4. The microorganism must then be reobtained from these inoculated
animals and cultured again.
Applicants have fulfilled Koch's law using pure, uncontaminated
cultures of cryptocides. Pure cultures were obtained repeatedly
from the various proliferative and neoplastic diseases of both men
and animals. Then they were injected into animals capable of being
infected. Gradually diseased areas developed which resembled those
from which the cultures were obtained. Then the pure cultures were
reisolated from the infected animals. Thus Koch's postulates were
A blood specimen of a terminal cancer patient was cultured, the
culture was extracted and the extract produced tumors in mice.
This demonstrates the growth factors.
In one attempt to produce antibodies and antiserum in sheep that
would be beneficial in the treatment of human cancer, sheep were
immunized with an attenuated or weakened culture. Twenty sheep
were examined and found to be free of disease. Some of the stock
cultures applicants had on hand such as cultures from human breast
cancer, from a sarcoma of a young boy, from a human leukemia, from
the Rous chicken tumor, from arthritis, and from fowl leukosis,
were attenuated. Applicants injected two sheep with each strain.
After about four weeks, some of the sheep became sick. Attenuated
vaccines from the cancer cultures were used weekly for
immunization. Several ewes aborted their young. The fetuses were
macerated. Some of the sheep developed very swollen painful joints
and could scarcely graze. Others looked poorly because emaciated.
Applicants realized that the vaccines which were attenuated were
still alive and had not fully immunized the sheep but had diseased
them. The sheep were bled in order to assay their serum for
antibodies. Sera was obtained but the sheep had to be destroyed.
Although the sheep had to be destroyed applicants learned that the
fowl leukosis serum agglutinated in high dilution the cultures
from the boy's sarcoma, that the breast cancer serum reacted with
the human leukemia isolates, and that the Rous sarcoma serum
reacted with all of the cultures. This meant that the cultures
from the human cross-reacted with one another strongly and with
the animal sera, showing that tumors are not tissue or species
Three chickens having fowl leukosos, a cancerous disease, and
which could no longer stand up, were taken to applicants'
laboratory and in a short time they were dead. Applicants made
cultures from their heart's blood. These grew to be the same kind
of cultures as those derived from all of the other tumors
experimented with by the applicants.
A pure, selectively grown bacterial culture of the type described
above, obtained from urine and blood using sterile precautions,
contains chemical substances related and/or identical to the
actinomycin group. To test this point, a phenolized pure culture
was acidified with HCl to pH 2 (Congo red/thymol blue) and was
left standing overnight after short boiling. A mixture of
n-butanol/conc. NaCl, equal parts by volume, containing a few
drops of glacial acetic acid, was used for extraction. After
gentle shaking for about 15 minutes a dark cherry-red layer of the
solvent was separated for further processing. This crude mixture
gave peak absorption at 440/450 mn and 410/425 mn values which
compare favorably except for a third absorption peak at 240 m.mu.
obtained by Waksman (Waksman, S. A., and Lechevalier, H. A.: The
Actinomycetes, vol. III, The Williams & Wilkins Co.,
Baltimore, 1962, p. 168) which was missing. From an ascending
paper chromatogram a reddish zone was eluted with ethylacetate and
an acetone-ether mixture, both gave upon evaporation some
microscopic crystals (red plates). The controls containing broth
and 2% phenol but no organism gave upon extraction a barely
yellow-tinted layer of the solvent. This exploratory separation
technique was then repeated with several 25-hour urine specimens
obtained from terminal cancer patients. After separation of the
organic layer from the urine specimens again a more or less
pronounced color was present ranging from dark honey-brown to
cherry red; controls taken from healthy persons did not contain
such colors. The presence of these dark colored compounds seems to
be most pronounced in terminal cases. All crude mixtures isolated
from cultures and/or urine were subjected to further separation by
METHODS FOR THE ISOLATION AND IDENTIFICATION OF A PLEOMORPHIC
INTERMITTENTLY ACID-FAST ORGANISM FROM NEOPLASTIC DISEASES, AND
THE PREPARATION OF CULTURES
Isolation from Urine (Crofton Method)
Obtain a midstream clean-catch specimen of urine in a sterlized
screw-top glass container.
Make up DiFco's brain-heart infusion agar: 37 grams of the agar
base are added to a liter of distilled water, heated to melt and
mix, and distributed into flasks or bottles of 95 ml amounts, and
autoclaved. Five percent (5%) human blood (outdated bloodbank
blood may be used) is added when the melted agar has cooled down
to 45-50 degrees C., and the mixture is poured into sterile Petri
Streak the surface of the blood agar plate with a sterile swab
which has been dipped in the urine. Incubate plate to 37 degrees
C. and examine after 24 hours. If growth has appeared, note types
of colonies, make duplicate smears, and stain one by Gram's stain
and the other by Alexander-Jackson's modified Ziehl-Neelsen
technique: flood side with Kinyoun's carbolfuchsin for 3 to 5
minutes in the cold, wash, decolorize briefly with 70% alcohol
containing 1 to 3% HCl as these organisms decolorize more readily
that M. tuberculosis, counter stain by flooding slide with
Loeffler's methylene blue and add 6 to 8 drops of normal (4%)
sodium hydroxide. Tilt slide to mix, and wash after 30 seconds.
APPEARANCE OF THE MICROORGANISMS ON BLOOD AGAR
Colonies--usual types of growth obtained
1. white discoidal, often hemolytic, and with a raised
center--having a fried egg appearance, but usually larger than
classic PPLO colonies grown on PPLO agar.
2. grayish muccoid, often confluent.
3. pigmented: yellowish, occasionally pinkish coral.
4. wrinkled intermediate SR worm-casting type resembling M.
5. dull granular surfaced, irregular edges, often hemolytic, and
resembling B. subtilis, but virulent for mice. Motile forms
transferred tp A-J broth produce a white or grayish white soft rim
or pellicle, and a toxin-like substance.
This organism tends to resist emulsification to some degree when a
loop of culture is rubbed with a drop of water on a glass slide to
make a smear.
The cancer isolate is either Gram positive or Gram variable. The
Gram stain is not the stain of choice, but should be used to
eliminate true Gram negative organisms, which show no Gram
positivity at all such as B. coli, proteus or pseudomonas.
Acid-fast forms may or may not be found, as this organism is
intermittently acid-fast rather than more consistently so as in M.
tuberculosis or other classic mycobacteria. However, if possible,
careful search for acid-fast forms is desirable, as they are
hallmarks of this mycobacterium-like organism. Slender filaments,
sometimes with lateral branching, and sometimes acid-fast, help to
distinguish it from common micrococci. The rods may be slender and
diphtheroidal, or thicker and subtillis-like. The latter sometimes
contain tiny brightly acid-fast bodies surrounded by a colorless
vacuole-like area or capsule. This appearance plus virulence for
mice and guinea-pigs distinguish them from subtillis rods. The
main morphologic forms are:
1. tiny acid-fast elementary-type bodies, often refractile;
2. Coccoidal forms of varying sizes with or without threads or
3. rods as described above; X, Y, and V forms commonly seen,
filaments may become very long and wide;
4. cyst-like bodies of various sizes from 3 to more than 10.mu.
and often containing smaller bodies which may be brightly
5. L or PPLO forms consisting of a lightly stained matrix
containing more deeply stained bodies of various sizes, or tangled
branching threads and ring-forms. These are revealed by
Alexander-Jackson's Triple Stain modification of the Ziehl-Neelsen
6. spore forms of oval shape seen in old cultures;
7. sub-microscopic bodies 20-70 m.mu. revealed by the electron
microscope, and of virus size.
The product of the growth of the specific Actinomycetales organism
is a chemical substance which is obtainable on recrystallization
(Wolter)--of a suspension of the isolate with conventional paper
chromatography annular separation procedures, and identifiable as
comprising a formation of
(a) red crystals identical with those described by Waksman as
having the absorption peaks indicated, supra;
(b) yellow crystals, similar to an actinomycin D fraction;
(c) formation of small placques of crystals similar in appearance
(d) a waxy, higher-alcohol formation, and
(e) a brownish, foul aromatic residue.
To an acidfied (pH 5) sample of urine in which the organisms are
grown, (phase I) and kept in the refrigerator, is added one-fourth
volume of n-butyl alcohol and the mixture is shaken for one-half
hour. The mixture is refrigerated until the layers separate.
Separation is done by decantation first and then by using a
separatory funnel. The butanol layer has attained a reddish brown
color and in some cases a yellowish color. This procedure results
in an aqueous phase (II) and a butanol phase (III). The aqueous
phase is extracted once or twice more in the same manner with
n-butanol, so long as the color appears in the extract.
A portion of the combined extracts is evaporated in an evaporator
at 35 DEG-40 DEG C. The dried residue is dissolved in a small
amount of methyl alcohol, solution (IV). Layers of a silica gel
preparation, MM-SGel-HR for thin layer chromatography, Machorey
Nagel and Co. 516 Durem, Germany, is spread on glass plated,
prepared, dried and stored in a dessicator. By means of a small
pipette, a spot of solution (IV) is placed on the silica gel layer
of one of these plates near one edge and near this spot at the
same distance from this edge and in the same way, another spot of
actinomycin D, (Merck, Sharp and Dohme), is placed in the same
manner. Both spots appeared bright yellow. Other pairs of spots of
these two solutions are placed on the same row using 5 m.mu., 10
m.mu. and 15 m.mu.. (One m.mu. equals one microliter.) Of the
solvents tried for developing the chromatograph, the most
effective was butanol-methanol-water in the ratios of volume of
6:1:3. 60 ml. of butanol, 10 ml methanol and 30 ml of distilled
water are mixed and put into a thin layer chromatography chamber.
The paper lining the walls of the chamber is wet by swirling the
solution in the chamber. Then the plate with the spots is placed
on edge in the chamber with the row of spots parallel to and near
the bottom but above the surface of the solution. A cover is
placed over the top of the chamber sealing it. The chamber is kept
in the dark during the process of separation since a better yield
elution is obtained. In previous runs each actinomycin D spot
travelled as a single spot leaving nothing in the pathway by
visible or ultraviolet light, and phase (III) left material spread
from the top streaking down along the pathways. However the top
was always at a level with and the same color as the actinomycin
D. Subsequently, after drying the plates, the actinomycin D spots
and those portions from the spots of phase (III) on a level with
the actinomycin D spots are cut out and separately eluted with
methanol, as are other portions from the spots of phase (III),
since these may contain other actinomycins than actinomycin D.
Visible ultraviolet and infrared absorption spectrograms are made
of the different elutes and compared. The TLC method is based and
adapted from methods reported by Cassani et al, J. Chrom. v. 13,
Blood and tissue cultures and urine samples obtained in accordance
with the foregoing procedures were extracted with and equal volume
of butanol, water, acetic acid (4:5:1) and the upper phase taken
to dryness. The residues were taken up into two 5 ml. portions of
ether and evaporated at 35 DEG-40 DEG C. Paper discs were dipped
into ether solutions of extract, the ether evaporated and the
discs placed on standard actinomycin assay plates. A standard
preparation of actinomycin D was also run. The zones of inhibition
are shown in Table I; zones above 15 mm. in diameter fall within
the standard range and are calculated in terms of actinomycin D
Act. D (ug/ml)
(1) 14 <0.2
(2) 15 0.2
(3) 16 0.4
(4) 12 <0.2
(5) 16 <0.4
(6) 14.5 <0.2
(7) NA <0.2
(8) NA <0.2
(9) 14 <0.2
(10) NA <0.2
(11) NA <0.2
(12) NA <0.2
(13) NA <0.2
(14) NA <0.2
(16) NA <0.2
(15) NA <0.2
(17) 22 4.2
(18) 16 0.35
(19) NA <0.2
(20) 18 0.8
(21) 17 0.5
(22) 13 <0.2
(23) NA <0.2
UV ABSORPTION CURVES
0.2 ml. of each ether solution was evaporated to dryness and the
residue taken up in 1 ml. of methanol and UV absorption curves
were recorded. End absorption at this level of purity prevented
measurement of 440 mm. absorption. Small peaks, typical of trace
amounts of actinomycin, were found with samples 1-6 inclusive and
sample 8 with a slight response present with sample 17. Definitive
biochemical tests such as the cytosine-guanine percentage of the
DNA have helped to classify the cryptocides microbial isolates.
Further concerning cryptocides, these organisms, which appear
primarily as small acid-fast granules in young cultures, and which
tend to become non-acid-fast in the larger forms present in older
cultures, may exhibit a number of morphologic phases with
intermediate transitional forms. These include (1) filterable and
submicroscopic bodies; (2) larger granules readily visible under
the light microscope an often resembling ordinary micrococci; (3)
larger globidal cystlike bodies and thin-walled sacs containing
the smaller forms; (4) PPLO or L type zoogleal symplasms without
cell walls; (5) rods of various sizes capable of developing a
characteristic motility; (6) long filaments and threads which may
show lateral branching; and (7) thick-walled spore-like bodies.
The lesions produced by these organisms in experimental animals
were generally pseudocaseous, degenerative in type, occasionally
neoplastic, and occurred principally in the liver, kidneys, and
lungs although at times, there was involvement of the heart,
spleen adrenal glands, stomach, lymph nodes, and omentum.
Dr. Afton Munk Livingston, and Dr. Virginia Livingston,
Transactions of the New York Academy of Sciences, May 1972, report
the recognition of the P. cryptocides organisms in the blood of
cancer patients compared with the blood of healthy individuals, of
which a summary follows. Examinations by darkfield microscope of
fresh blood, and also by brightfield microscope using supravital
stains serving as a diagnostic and prognostic tool in following
the course of the cancerous disease in the patient in conjunction
with several other microbiological evaluations.
PREPARATION OF SLIDES FOR BLOOD EXAMINATION
The patient's finger is immersed in 70 percent alcohol and air
dried. A steril lancet is used to puncture the finger, a small
drop of free-flowing blood is placed on a sterile clean slide and
covered with a sterile covership. Care is taken that the blood
does not flow beyond the edge of the coverslip. Using a small
weight for approximately one minute, light pressure is applied to
the coverslip to spread and separate the blood cells. The
preparation is then examined under darkfield at .times.750 and
.times.1350 magnification. For lightfield examination, the same
method is followed and in addition, a small drop of 1 percent
aqueous sterile crystal violet, freshly prepared and filtered,
gently applied to the preparation. If the number of organisms, to
the blood as well as the motility of the various stages are to be
evaluted, then the blood is diluted 1:100 with sterile distilled
water using a sterile red-blood-cell diluting pipette. The pipette
is then thoroughly shaken and a few drops are expelled from the
pipette into a sterile Petri dish. A small measured amount of 1
percent aqueous crystal violet is added. This mixture may then be
used to flood a blood counting chamber. This method provides a
quantitative estimate of the numbers of the organism as well as
their motility, which may last as long as fifteen minutes.
However, for the usual brightfield examination of the blood with
crystal violet, the blood drop is placed directly by the slide and
the small amounts of crystal violet is added before the coverslip
is placed over the preparation and light pressure applied.
DARKFIELD EXAMINATION OF UNSTAINED FRESH BLOOD PREPARATION
A number of interesting observations may now be made by the
darkfield, pulsating orange bodies in the red cells may be
observed. In the background, there are bright dancing forms which
appear to be small L-forms of the organism. In several infected
hosts a number of motile rods may be observed. Spheroplasts and
mesosomes both large and small are present. These may have many
fine delicate vibrating forms in their periphery. Forms resembling
a medusa or a octopus with waving filaments may be present.
Organisms may bud from the surface of the red cells and from fine
hairlike filaments which resemble the handle of a tennis racquet.
There may also be numerous threadlike filaments free in the serum,
varying in size, some 10-15 microns in length. These are motile
and appear to wind in and out around the red cells. There are also
long tubular structures 50 microns or more in length, and about 10
microns in width that are milky white, highly luminescent,
containing numerous refractile granules. The tube in some cases
appears to arise from a coalescence of the L-forms or to bud from
a spheroplast. It is transparent since cells can be seen through
it. When the tube wall disintegrates the refractile bodies are
released in the serum and may enter fresh red cells. There are
also large round milky white forms appearing to be protoplasts
about 20 to 60 microns in diameter which contain granules
resembling spheroplasts or mesosomes. The protoplasts may have
budding forms at the periphery and may release rather large
vesicular refractile bodies resembling the spheroplasts or
mesosomes. At times, the extruded mesosomes are large enough to be
mistaken for red blood cells, but they do not have the bluish
tinge of red cells seen in darkfield. Rather, minute dancing
particles may later appear within them.
In addition, shrunked red cells with a ground-glass appearance
spiculated at the periphery may be observed. We have termed these
structures "spent cells" since they appear to be red cells that
have been consumed by the parasites. They are lighter and smaller
than normal erythocytes and have a tendency to be pushed to the
periphery of the blood drop when it is prepared for examination.
Changes in the character of the leukocytes are also apparent. Many
leukocytes in the advanced stages of diseases appear smudged,
inactive and only dimly luminescent whereas normal leukocytes have
vigorously active granules and active amoeboid movements. Under
some circumstances great numbers of fine spicules occur in the
dark field. These are very delicate and appear to arise from
minute L-forms. They are not thrombocytes. At times they appear to
shed from the surface of the protoplasts. Why they should be more
numerous at one time than at another is not understood but their
appearance may be related to the pH of the blood. Orange
crystalline forms of the organisms as well as free crystals may
also be seen in and around the microbial clusters in the plasma.
They apparently arise from the waxy-secretions of these
mycobacteriumlike organisms. These are the crystals that have been
extracted from pure cultures and urines of terminal cancer
patients and that have been used for various types of bioassay.
BRIGHTFIELD EXAMINATION OF SUPRAVITALLY STAINED FRESH BLOOD
On a blood preparation stained with crystal violet and examined by
the brightfield method a clear white light and a magnification of
at least .times.1000 microbial forms are revealed that are not
seen in the darkfield. There are large branching fungal forms that
are not luminescent in the darkfield. These fungal forms may
extend over a considerable area involving several microscopic
fields. Some of these are branching and appear to have conidial or
frutting bodies attached to their branches. Microcolonies may be
clearly seen surrounding individual red cells and some appear to
arise from parasites extruded from the cells. These microcolonies
appear to develop into a network of interlacing branching fungal
filaments which act as bridges between the red cells and cause
them to adhere in clumps. The number of fungal forms which hold
the erythrocytes together or adhere to their surfaces may be
directly related to the sedimentation rate. The greater the
adherence of the erythrocytes due to the mycelial forms, the more
rapid the sedimentation rate. The red cells become separate and
free as the number of both intra-and extracellular parasites
diminish. The stained preparations in the counting chamber have
L-forms, which appear much more numerous than in the darkfield,
and occur in clusters, which have marked Brownian movement. These
clusters agglutinate and become motionless after ten to fifteen
minutes. Introduction of gamma globulin or specific antiserum
under the coverslip of the counting chamber caused instant
agglutination and cessation of motion. By this method, antibody
activity of blood serum can be roughly estimated. Other dyed
microbial forms in the brightfield may be compared with those in
the darkfield. The vibrating orange bodies in erythrocytes in the
darkfield appear as violet bodies in lightfield. The brightly
luminous tubles take on a light violet color with deep purple
The same comparison between darkfield and stained brightfield
preparations may be drawn by examining blood cultures grown in
broth. Hanging drops of cultures sealed with sterile vaseline are
preferable to ordinary wet perparations since they are safer to
handle and can be preserved for a longer period of time.
Conventional staining of slide preparations appears to break up
many of the delicate microcolonies and interlacing fungal forms.
Wet supravitally stained preparations in hanging drops also
indicate the degree of motility of many of the microorganisms.
Other dyes have been used which penetrate to some extent but do
not provide sufficient contrast. They are Sudan black, saffron
yellow, Congo red, May Grunwald, toluidine blue, gentian violet,
as well as several others.
All cancerous patients yielded L-forms as well as other
pleomorphic stageson blood culture which, on further cultivation,
developed the typical acid-fastness of the Progenitor cryptocides
group as previously described. However, the cancer patient even in
the advanced stages of the disease is usually afebrile. Comparable
numbers of microorganisms other than the Progenitor cryptocides
groups might be expected to produce an acute febrile reaction.
There undoubtedly can be a mild or transitory bacteremia in blood
due to relatively nonpathogenic bacteria such as some of the
diphtheroids. However, with the previously described methods, the
great numbers of the Progenitor group as a silent but lethal
bloodstream infection may be readily demonstrated. Advancing
infection of the bloodstream with P. cryptocides is relatively
asymptomatic until large numbers of the organisms are present and
there is a concomitant breakdown of the immunological and
The autogeneous vaccine is known to exist by disc-saturated
inhibition on culture plates of extracts and also from the serum
of cancer patients.
Administration of the autogenous vaccine should be initiated by
high dilutions of the lowest order of dosage at twice weekly
intervals, with gradually increasing dosages until overdosage
symptoms occur. Preferably, subcutaneous injection of the
autogenous vaccine in a suitable pharmaceutical carrier, such as
sterile water or saline solution may be employed, although oral
administration of the product in a suitable carrier also may be
Use of the autogenous vaccine of this invention may prove to be of
value in the palliative treatment of animals and humans afflicted
with various forms of neoplastic diseases, as indicated by
treatment thereof with autogeneous vaccines made from a suspension
of the isolates in 2% phenol. In preparation, the vaccine is
allowed to stand overnight at room temperatures, centrifuged and
further diluted with 0.5 phenol or saline.
Subcutaneous injection is initiated with the highest dilution of 1
million organisms per ml and 0.1 ml twice weekly, until overdosage
symptoms occur. Therapy is continued with higher concentrations,
e.g., 10 million and 100 million organisms/ml. Oral administration
of the same dosage can also be employed.
A study of one hundred random blood samples, taken in the office
of a physician who specialized in allergy and immunologic disease,
showed that all tumor-bearing patients, in comparison to office
personnel used as controls, gave positive cultures for the
cryptocides organism. A number of patients with chronic
degenerative disease were also positive. While many patients who
had reached a healthy old age were negative, several "tired" young
people without apparent disease were positive.
A reddish brown material has been extracted from the tissue, urine
and blood of cancer patients in increasing amounts as they became
terminally ill, and (this material has not been found in normal
controls. It is carcinogenic for mice, increasing the incidence of
pulmonary tumors. The biological effects have been assayed in
preliminary studies with tissue-culture systems and with
tumor-genesis in mice.
The applicants have found the presence of actinomycin-like
crystals in body tissues and in cultures.
The basic requirement for formation of the cancer cell is the
causative microorganism; all other factors such as coal-tar
irritants, other microorganisms, the aging process, any chronic
irritants leading to poor local resistance and giving rise to
immature, succeptible reparative cells, may prepare the living
matter, e.g., for the multiplication of the cancer organism and
its penetration into the cyptoplasm and nucleus of the host cell.
Apparently the organism cryptocides can invade both cytoplasm and
nucleus of host cells in any type of host tissue when body
defenses are lowered. In experimental animals it can cause lesions
that appear as necrotic abscesses, granulomas, fluid-filled cysts
of neoplasms. The type of lesion apparently depends on specific
and nonspecific immunocompetence and the age of the host.
Certain chemicals can have a tremendous effect upon the entire
hormonal system. One of these substances is actinomycin produced
by several of the actinomyces organisms and probably by many of
the Actinomycetales. There is a whole array of chemicals and
biologic produced by this group of microbes, which have been used
as antibiotics and antineoplastic agents in some cases. The
actinomycins even in very high dilution of one part in a billion
or more may have a profound effect upon the entire business of
life. The important thing to remember is that no funtcion of the
body is exempt from this toxic material which is produced by these
microorganisms belonging to the Actinomycetales. Not only are the
normal functions of the host's hormonal system deranged but there
are "false or counterfeit hormones" produced with further throw
the body off balance. There is a practice of castrating men and
women to arrest the growth of cancer. If castration is successful
in prolonging life, then adrenalectomy and pituary gland removal
might be done when the effects of tumor inhibition from the
castration have worn off. This hormonal ablation presents a grim
picture to say the least. Applicants believe that the hormonal
stimulation of the sex glands, the adrenals and the pituitary are
the result of toxic materials, hormonal derangers and counterfeit
hormones, such as, phytosterols produced by the Cyptocides, that
upset the balance of the patient's hormones not only by inhibitory
effects by production of pseudo or counterfeit hormones that act
on the physiologically controlled, normal glands causing abnormal
response. Also various kinds of cell poisons and inhibitors
destroy the efficacy of the lymphocytes to attack the cancer
cells. The cancer cells themselves are prevented from reaching
maturation by these cell poisons They are sick cells unable to
reach a normal maturity and normal function, whereever they are
located and whatever tissue they may be whether glandular,
interstital, bone or blood.
The most important thing is to try to destroy the microbes that
were producing the aberrant cell inhibitors and false hormones.
However, it has been reported that low testosterone levels have
been induced in patients with cancer of the prostate by treatment
with diethylstilbestrol, a synthetic hormone; and
amino-glutethimide, a powerful inhibitor of adrenal corticosteroid
biosynthesis, with patient improvement. Furthermore, an
immunological mechanism appears to be involved; the inhibition of
steroid biosynthesis. By removing the lympholytic effect of
corticosteroids, there is produced a marked hyperplasia and
increase in the number of circulating lymphocytes which potentiate
the immune response. The presence of lymphocytotoxic antibodies
has been reported in patients with prostatic cancer. Perhaps this
steroid is a "false steroid" and antagonism by the
amino-glutethimidine and diethylstilbestrol may permit an increase
in the production and circulation of normal lymphocytes capable of
attacking the cancer cells.
The role of steroids in chronic diseases was demonstrated by
Edward Kendall and Philipp Hench in their studies in rheumatology
for which they received the Nobel Prize in 1950. It is true that
the steroids do not have an inhibitory effect on these diseases
but at the expense of suppressing immunity and permitting the
underlying latent infection to continue or to increase in its
It is known that a bacterium belonging to the Actinomycetales was
able to produce unlimited amounts of steroids from the Mexican
It is stated that some steroids decrease the numbers of
circulating lymphocytes as well as blocking immunocompetence.
Perhaps the "false steroids" are really responsible for this
action. It has been shown that certain toxic antigens prevent the
lymphocyte from maturing and become immunocompetent. Leukemia, or
an accumulation of large numbers of cells, either lymphocytes or
polymorphonuclear leukocytes, may represent a blocking of the
pathway to maturity by a toxic agent such as a steroidal or
actinomycinlike compound produced by the cryptocides. Perhaps the
blocking factor may be related to a protective mechanism directed
toward making the cryptocides insusceptible through some
biochemical fraction that blocks the immune reaction of the
The present invention is useful in the treatment of man and/or
animal. Safety and effectiveness of the present invention has been
demonstrated in animals and has been indicated in the treatment of
humans by administration of the aforesaid vaccine form.
Among the various neoplastic diseases (often termed diseases of
unknown etiology) subject to palliative treatment are cancer,
tumor of the lymphoid tissues, Hodgkin's disease,
reticulo-endothelial tissues, arthritis, lymphosarcoma, the broad
spectrum of epidermoid cancer, scleroderma, adenocarcinoma,
fibrosarcoma, liposarcoma, myosarcoma, acute glomerulonephritis,
leimoya sarcoma, osteogenic sarcoma, chondro sarcoma, myeloma,
rous chicken sarcoma, coal-tar-induced cancer, fowl leukosis,
animal tumors such as Rous, Walker, Sprague-Dawley, Shope and
Sarcoma 180, and the like. Many of the foregoing are degenerative
and antoimmune diseases.
Malignancy is a neoplastic infection, which depends on the number
and virulence of the invading organism, the susceptibility of
various organs to it, as well as the natural immunologic
components of the host.
The reddish-brown crystalline substances extracted from broths
containing organisms within this invention are antibiotic to the
bacteria and to all strains of the producing organisms themselves.
The preferred media for growing cryptocides are obtained with
Alexander-Jackson's broth, and with Wuerthele-Caspe's autoclaved
chick embryo agar. The method of preparation of these two media is
ALEXANDER-JACKSON'S (A-J) SENSITIVE PEPTONE BROTH
beef lung, cut up
peptones 20 grams; 5 grams of each of
(a) myosate, (b) gelysate,
(c) trypticase, (d) phytone
glucose 10 grams
glycerol 80 ml
Boil the beef lung and water for 30 minutes. Filter through cotton
or very coarse paper into a flask containing the other
ingredients, and heat to dissolve. This crude lung broth can be
autoclaved and stored in the icebox, and clarified subsequently.
Autoclaving for a second time does not seem to produce any adverse
A 1 to 2 mm layer of infusorial earth (Standard Filter Cel of
Johns-Manville Co.) is deposited on a No. 42 Whatman paper disc by
laying the disc on a Buchner funnel, applying suction, and then
carefully pouring on about 500 ml of a 5 percent suspension of
Filter Cel. After the deposition of the layer, when the water goes
through clear, the suction flask is well rinsed out. The hot
medium can now be filtered through the prepared disc into the
The medium should be filtered a second time through a Buchner-type
funnel with a fine fritted glass disc, or else passed once more
through the same Filter Cel.
The pH of the medium should be adjusted to 7.4 with sodium
hydroxide. The medium is then tubed into screw-top glass tubes
150.times.25 mm (Kimble Glass Co., Toledo, Ohio). The tops of the
tubes are not screwed tightly until after autoclaving. Autoclave
for 15 minutes at 15 pounds pressure. Place about 5 ml of medium
into each tube; or place 50 ml in a 250-ml Erlenmeyer flask for
primary isolates. Close the flasks with cotton plugs held in a
single layer of gauze, and protect the plug by a paper drinking
cup or cone.
The A-J broth is obtainable from the Colorado Serum Company of
Whole fresh citrated or untreated blood, 0.2 ml is added to 2 ml
of broth at pH 7.4, and incubated for a week. A transfer to fresh
broth is then made to rid the culture of antibodies or other
inhibitory substances. After several days, the organisms appear as
a mat at the bottom of the tube. When motile rod forms are
present, a soft while ring or pellicle appears. Growth is often
seen climbing up the side of the glass tube.
The peptones included in the above broth have been studied
individually. Myosate, a pancreatic hydrolysate of heart muscle,
favors small virus-like and coccoidal forms. Gelysate, a gelatin
hydrolysate, appears to favor slender acid-fast rods and
non-acid-fast rods containing acid-fast granules. Phytone a papaic
digest of soya meal, and trypticase, a pancreatic digest of
casein, both favor the readily growing motile rods. A combination
of all four peptones provides a medium which allows a wide variety
of forms to develop, and makes it easier to recognize the presence
of the organism in primary isolations.
WUERTHELE-CASPE'S CHICK EMBRYO AGAR
The contents of 8-15 days old embryonated hens' eggs are ground up
on a Waring blendor, mixed with 1.5 percent melted agar, tubed in
screw-top glass tubes, preferably large ones, slanted, an
autoclaved at 15 pound for 20 minutes in the slanted position.
Examples of other media which can be used are Difco's brain-heart
broth with and without glycerin, Dubos medium, Alexander-Jackson's
modification of von Szabocky's glycerol lung broth, dextrose blood
agar, Alexander-Jackson's adaptation of Bushnell's poi agar,
Petragnani, Lowenstein-Jensen, Dorset egg media, Witte's peptone,
Difco's bactopeptone, Armour's peptone, a Merck peptone and
METHODS OF IDENTIFYING CRYPTOCIDES
This invention also involves a test that will show the existance
of a neoplastic disease before the tumor exists, or the existance
of a chronic underlying infection in man and/or animal. Until now
any aberrant symptom of a patient has to be evaluated in the light
of a latent cancer until it was ruled out. A fever of unknown
origin could turn out to be a sarcoma somewhere in the body made
manifest weeks later after much laboratory work and X-rays. By
then, it was already too late, to do anything. Even if it had been
known that cancer was imminent there was no treatment. There was
nothing to do but wait until a tumor presented itself and then
attempt to cut it out or destroy it by radiation or chemicals.
(Applicants' invention involves a cure for such cancer.)
Tests for determining the presence of cancer such as the Pap
smears testshave serious problems associated therewith. In the Pap
smears tests, the body cells that are cast off from the uterus,
cervix and vagina are smeared from the cervix, are placed on a
silde and stained. Not only is the presence of cancer cells
detected but the amount of estrogen in the body is indicated by
the size and shape of the nucleus of the cell in relation to the
cytoplasm. This test is useful in determining the stage of
menopause in women. Unfortunately, when the smear for cancer is
positive, the cancer is already there. However, it does permit
early detection of some kinds of cancer of the female reproductive
organs. The same method of cell determination is now applied to a
number of other sites such as lung and stomach.
As cancer is an infection, surgery, radiation and chemicals cannot
eradicate a continuing infectuous process. For example, cobalt
machines may reduce the size of tumors but contribute very little
to the long-term cure of the disease.
The test of this invention allows a screening program of the
entire poputation by means of routine blood cultures to determine
the presence of the cryptocides bacteria correlated with
evaluation of blood smears and related to immune competency by
various methods of antigen-antibody determination.
There are a number of identifying biochemical tests that can be
applied but these are too time-consuming and expensive for a
routine laboratory. In the dying patient, a few drops of blood
taken from the antecubital vein of the arm will grow out furiously
on direct plating on the solid blood plates. Usually, isolation
from blood is done by placing ony a few drops of blood, about
five, in the bottom of a peptone-broth tube, and incubating. The
organisms can be readily recognized either in hanging drops of the
living cultures or by appropriate staining. The organisms grow up
the side of the tube forming a lacy pattern and then produce a
pellicle or doily on the surface. These usually signify the
presence of motile rods. This is a good stage from which to make a
vaccine. As the pellicle ages it has a tendency to drop into the
tube again the spore stages are then formed. The spores cannot be
used for vaccines as it is almost impossible to kill them. The
liquid cultures will often transfer to solid media plates.
Applicants' sensitive peptone broth for primary isolation is
useful, and it can be obtained from the Colorado Serum Company in
Denver, Colo. Dr. Diller's paper gives the various methods of
isolation using the technique of Von Brehmer, Glover, Seibert and
others. Applicants have also used synthetic broh media for primary
isolation but these proved to be too toxic on animal
There are several other ways of making primary isolations of the
cryptocides. Sterilely obtained tumor tissue fresh from the
operating room can be placed into liquid media and later
transferred to solid blood plates. Some people have ground up the
tumors, filtered them and then cultured them. This is difficult
because of the problem of maintaining sterility. These methods led
to the recovery of the specific microorganism, the cryptocides.
Still others have made various extractives of the tumors with
alcohol, acetone or other solvents and used these for the vaccine.
Another method is to grow the organism from one of its favored
spots, the roots of infected teeth or tonsils. However, the mouth
contaminants must be eliminated. Still another way is to dilute
the patient's blood with equal parts of distilled water in order
to disrupt the red cells wherein the parasites are contained as
well as in the serum. The tubes are lightly boiled over an open
flame two or three times and then incubated for eight to twelve
days. Intervening examination of the blood will reveal the rate of
growth. When the growth is abundant, usually in ten to twelve
days, the blood can be filtered to remove larger particles, then
formalinized, standardized and tested for viability. This method
may have some advantages over the whole-cell antigens obtained by
the Crofton method, because the whole-blood cultures will also
contain toxins and antitoxins as well as many of the minute forms
which do not grow out on artificial media. This is the German
The following is a description of applicants' test method for the
quick detection of the presence of the chorionic gonadotropic
hormone (which is produced by cryptocides) in urine which
indicates the existence of a chronic underlying infection in
tissue and blood which produces the chorionic gonadotropic
hormone, thus eliminating the need for study of individual
colonies. This test can be of great importance to the medical
profession, particularily in determining the presence of cancer or
the likelihood of cancer proliferation. The test can be used in
man and animal.
Applicants theorize that the abnormal production of the steroid
chorionic gonadotropic hormone, keeps the cancer cells growing.
Chorionic gonadotropic hormone is made by the microbe Cryptocides.
This tool allows the diagnosis and prognosis of chronic underlying
infection or condition. Treatment can follow.
Describing the diagnoistic test, the way that applicants have
found to indicate whether or not choronic gonadotropic hormone is
present in the urine is: to take a predetermined amount (usually
10 ml.) from a urine specimen of the person or animal; prepare a
blood culture using the urine sample; allow the blood culture to
set (e.g., one day); take a predetermined amount (e.g., 3 to 5
drops) from the blood culture and directly place it in the peptone
broth described elsewhere herein; and incubate (e.g., at 37 DEG C.
for 24 hours) the peptone broth. Blood plasma or serum can be used
in a suitable container such as a test tube. Blood is its own
media and no media need be added to it. This test procedure will
give a negative or positive result depending upon the absence or
presence, respectively, of chorionic gonadotropic hormone in the
urine speciment. (It should be noted that minimal essentially,
undetectable amounts of chorionic gonadotropic hormone may be
present in the urine due to the presence of pathologically
insignificant amounts of Cryptocides that are universally present
in man and animal.) A positive indication (colony growth) will
occur, if it is going to at all, in approximately ten days.
(The microorganism from the colonies from the urine-blood cultures
may be tumorogenic and antibiotic.)
Negative, e.g., hormone, indications are always obtained when the
sample from the blood specimen is first cultivated on a plate
(e.g., agar) and then the colony is placed in the peptone broth.
Negative indications are always obtained when the urine is culture
on a blood plate (i.e., media has been added) and the colony is
then placed (e.g., place blood plate with colony between fingers
and squeeze over top of the broth test tube) in the peptone broth.
In both instances negative indications are present even when more
than normal amounts of chorionic gonedotropic hormone are present
in the urine. Proper postivie indications are obtained when blood
in test tube is used, but improper negative indications are
obtained when blood in plates using a solid media is used. It has
been reported that Cryptocides is anaerobic, which may explain the
aforegoing. Also there may be present a mixture of phases of the
microbe due to the pleomorphic nature of microorganism.
A positive indication means that the person or animal from whom
the urine specimen was taken has a chronic underlying disease or
infection, such as, cancer. This test can be used to detect all of
the above listed neoplastic diseases.
Describing the prognostic test, the same procedure is used as
described above for the preparation of the incubated peptone
broth. Any resultant positive indication is measured
conventionally for intensity or magnitude and/or length of
duration of such positive indication. This gives results or
measurements that can be compared with prior or future results or
measurements from other incubated peptone broths made from the
same patient. The prognostic test gives qualitative and
Since the presence of the bacterium can be proven by the product
it makes, if the patient is well, the bacterium is attenuated
(surpressed) and does not make significant amounts of the product.
When the patient gets worse, cryptocides makes more chorionic
gonadotropic hormone so prognostic tests can successively be made.
Corroboration was achieved by passing cultures, having positive
indications, from primary blood isolates into the next test tube
with the presence of blood. The passed cultures were allowed to
grow until visible growth was seen. The growth was tested and it
gave positive indications.
Another test for determining or detecting the presence of
chorionic gonadotropic hormone which indicates the presence of a
chronic underlying disease or infection in blood and tissue which
produces the chorionic gonedotropin hormone involves a skin test.
The vaccine described above is subcutaneously placed under the
skin. If a welt appears, it is a positive indication that
chorionic gonadotropin hormone is present and that a chronic
underlying disease or infection exists in the patient.
A variation of the above skin test involves conventionally
extracting a tuberculin-like substance from the vaccine described
above, and subcutaneously inserting the extracted substance. If a
welt appears, it is a positive indication that chronic
gonadotropic hormone is present and that a chronic underlying
disease or infection exists in the patient.
The urine test is the preferred detection test.
Applicants' urine test is exquisitively sensitive to a chorionic
gonadotropic hormone--there are no interferring substance In other
words, applicants' urine test is specific to chorionic
gonadotropic hormones--it is a serological, higly specific test.
The urine speciments from seven cancer patient were each placed
into separate test tubes containing only A-J peptone broth as the
medium. There was specific growth in each test tube. The colonies
were removed in each instance and were subjected to separation
(extraction) processes until only the microorganism was left. Each
of the seven isolated microorganism specimens were placed in
separate test tubes containing only A-J peptone broth (no blood or
agar was used). In each of the seven tubes, a positive reaction
was obtained confirming the fact that the microorganism which
produces chorionic gonadotropic hormone was present.
Applicants do not known of any other microorganisms besides
Cryptocides which produces chorionic gonadotropic hormone, which
gives a positive result in applicants' urine test.
It is known that the standard pregnancy test will give a positive
indication when chorionic gonadotropic hormone is present. It is
also known that a positive indication can be obtained in the
pregancy test from aspirin.
Some state that cancer is essentially a cesspool for the
collection of microorganisms, but applicants have found that only
Cryptocides causes cancer and it is the only microorganism which
produces chorionic gonadotropic hormone. Dr. Ross and others have
stated in several instances that chorionic gonadotropic hormones
are produced when a patient has cancer and that the chorionic
gonadotropic hormone emanates from cancer.
Applicants have found that the chorionic gonadotropic hormone
and/or biologically related hormones and sterols which yield a
positive indication in a standard pregnency test and which can be
identified by gas chromatography emanates from Cryptocides, which
causes the cancer. An example of such a biologically related
sterol are the phytosterols. An example of such a pregnency test
is Walpole's "UCG-Test" pregnancy test which detects human
chorionic gonadotropic immunologically in the urine of pregnant
Dr. Ross's tests used blood and found the presence of chorionic
gonadotropic hormone. These are called the L-tests. Applicants are
able to detect cancer by detecting the presence of chorionic
gonadotropic hormone in urine. Applicants' can use their test in a
diagnostic sense, that is, it can be used to determine if cancer
is present. Applicants' can also use their urine test in a
prognostic sense, that is, they can use it to see whether or not
the amount of chorionic gonadotropic hormone has increased or
decreased, and can use it to indicate the treatment to be used
(depending on whether the patient's condition is better or worse).
Lactrile states that the chorionic gonadotropic hormone which is
in the embryo stage is different from the other stages.
Applicants' urine test detects chorionic gonadotropic hormone
regardless of the stage it is in. (Kitts stated that all of the
cancer cells are in the first stage).
Chorionic gonadotropic hormone is a water soluble,
Researchers for a number of decades have been seeking tumor
antagonists not only in the form of antibiotics, which are
chemicals secreted by other microorganisms, but through potential
immunization by the use of other microorganisms. These biologicals
are in sharp contrast to the chemotherapeutic agents that seek to
destroy the dividing tumor cell regardless of the entailed immune
suppress Applicants have used the causative agent itself as a
menas of immunization.
Applicants have prepared and used an autogenous vaccine for the
treatment of chronic, ongoing infections. Customarily the vaccines
are prepared from urine, nasal, throat and bowel secretion as well
as from various tissues and other secretions. The vaccines are
used for the building up of immunity in the chronically ill
patient who suffers from a failure to produce immune bodies
against his chronic infection. This state of nonresponse is called
immunoincompetence. Applicants do not represent to the cancer
patient that the use of autogenous vaccine is proposed for the
treatment of cancer but for their underlying failure of immune
competence. In many cancer patients applicants do not use
autogenous vaccine. The use of vaccines must be carefully weighed
in the evaluation of the patients' immune status. In some cases
the use of vaccines are actually contraindicated. In the seriously
ill cancer patient the most important thing is to raise the
patient's immunity by the use of fresh whole blood transfusions
from suitable donors, and by the use of antibodies such as gamma
globulin. The next most important thing is to treat their chronic
underlying infection whatever it may be with suitable antibiotics.
The removalof harmful substances from the diet is essential as
well as the addition of needed vitamins and nutriments that may be
lacking in the seriously ill because of lack of appetite and
weight loss and faulty diet. Applicants do not believe that
vaccines can cure the cancer patient. It is one of the modalities
used for the chronically ill whatever their disease in the effort
to restore their resistance to an ongoing; underlying chronic
It is now incontrovertible that the cancer disease results in the
loss of immunity yet it is treated with radiation which destroys
immunity and with drugs which encourage cancerous growth. An
abstract of "Carcinogenicity Studies of Clinically used Anticancer
Agents", D. P. Griswold, J. D. Prejean, A. E. Casey, J. H.
Weisburger, E. K. Weisburger, H. B. Wood, Jr., and H. L. Falk.
Southern Research Institute, Memorial Institute of Pathology and
Baptist Medical Center, Birmingham, Ala. 35205, and National
Institutes of Health, Bethesda, Md. 20014, shows a number of
drugs, which produce cancer in experimental animals, yet whose use
is advocated by a government agency and the medical profession.
Such drugs include Melphalan, Chlorambucil, uracil mustard,
Natulan, dimethyltriazenoimidazole carboxamide, and 1,3-bis
(2-chloroethyl-) and 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea.
Some of these are so-called anticancer drugs. Tumors of several
types and at a variety of sites were seen in mice and rats during
a testing of such drugs for carcinogencity.
Applicants repeat that they do not treat cancer with vaccines, use
autogenous vaccines obtained from the patients' own tissues and
body fluids to treat an underlying chronic infection. These
organisms are not present in cancer alone but also in a host of
collagen diseases and in healthy carriers as well, and that the
use of vaccines for chronic disease states is an accepted modality
in therepeutic medicine. Use of applicants' autogenic vaccine is
therefor beneficial to a patient whether or not the patient has
cancer. However, by giving the patients good nutrition, by helping
them fight off chronic infection, and by using any and all
accepted modalities that may be helpful, applicants assist the
patients in throwing off their diseased condition, whatever it may
The production of vaccines from blood cultures is a rather long
and tedious procedure. Applicants use the following method for
making autogenous vaccines from urine. Applicants use sheep cell
blood agar with phenyl ethyl alcohol, which inhibits the growth of
E. coli, a common contaminant. Either the patient leaves the urine
as directed with a testor or follows these directions at home:
1. Boil a screw-top bottle and top for twenty minutes. Let it
cool. Remove with sterile tongs which have been boiled or
disinfected in rubbing alcohol.
2. Take a bath. If a female, take a douche and wash off
3. Using three balls of sterile cotton wash off from front to back
three times over the perineum with either Phisohex or some other
mild disinfectant such as St 37. If a male, pull back the foreskin
and wash throughly three times separately with each of the three
4. Start the urine stream over the toilet bowl and then catch the
midstream into the sterile bottle without contaminating the
inside. Be careful to keep the fingers out of the inside of the
screwtop cap. Tighten the cap throughly to prevent leakage.
When the urine is received in the laboratory it is streaked onto
the sterile blood plates using sterile swabs. The plates are then
incubated in the usual way. Generally within 12 hours small
colonies have formed. When these have been properly identified by
allowing the growth to continue for a day or two to be sure that
the characteristic colonies are present, then a single colony is
studied by Ziehl-Neelsen stain, Kinyoun type, for acid fastness
and the characteristic morphology. Then a single, identified
colony is spread on one or two additional plates where they are
incubated until sufficient growth has accurred. Stained
preparations are again examined. The colonies are then swabbed off
into a 2 percent phenol solution and permitted to stand overnight.
Then after about eight hours, the phenolized cultures are diluted
to 0.5 percent phenol. It requires about two weeks to complete the
sterility tests and to make several dilutions according to
government regulations. Autogenous vaccines tailored for each
individual are prepared, but this procedure is not so limited. The
vaccines are made up into 10 million, 100 million, and 1,000
million (=1 billion) organisms per c.c. The lowest amount, 10
million organisms per cc, is used as the starting bottle for
progressive immunization. Doses are taken every three to five days
depending on the reaction. It is wise to start with 0.1 cc by
subcutaneous injection of the lowest amount and observe for
evidence of redness or soreness at the site of the injection or
symptoms of hpersensitivity such as mild fever, lalaise, or muscle
or joint pains. if there is a mild reaction, the patient waits
until it subsides. before repeating the same dose or smaller in
three to five days by mouth. If there is no reaction, then the
dose is increased by 0.1 cc to 0.2 cc and administered first by
subcutaneous and then orally in three to five days. The third
week, the dose is increased again by 0.1 cc to 0.3 cc
subcutaneously and repeated orally again in three to five days.
The oral dose is taken under the tongue and held in the mouth for
absorption. The vaccine is increased in this manner until twenty
drops are taken. Then the next higher dilution of organisms is
started in bottle number two or 100 million organisms per cc. The
starting dose is only 0.1 cc since the second dilution is ten
times as strong as the first one. Again the doses are increased
gradually and so on with the other bottles of the vaccines. A
vaccine usually lasts six months but if there is quite a change in
the character of the organisms under treatment the occasionally it
is good to prepare a new vaccine in three months. This method just
described is applicants' preferred method of preparation of
autogenous vaccines and their administration.
The single most important factor in the presentation of vaccines
is to rule out common contaminents. The colonies can be entirely
confluent in severely infected hosts so that a transplant must be
made in order to isolate individual colonies for study. The
typical colony has an umbonate (fried egg) shape and may or may
not be hemolytic. The colonies may also be wrinkled or smooth,
china white or pale tan and even pale pink or orange when grown in
the dark. The slides are made by lightly wiping a culture from the
plate with a sterile cotton swab or with a platinum loop onto the
surface of the glass slide and fixing it with gentle heat. One
colony only should be selected and a cross-section should be
studied by taking samples from the center outward to the periphery
to obtain the different pleomorphic stages. If the material is
handled gently they ray formation of the growth will not be broken
up. The Kinyoum modification of the Ziehl-Neelsen stain is used
since it can be applied in the cold for five minutes and does not
require heating. The red dye is washed off with sterile distilled
water and the slide is then briefly decolorized with 1 percent
hydrochloric acid in 70 percent alcohol. The Cryptocides organisms
are more sensitive to decolorization by acid-alcohol than the
tubercle bacillus. The slide is washed again and the methylene
blue counterstain is applied, 6 to 8 drops of normal (4%) of
sodium hydroxide are added. After 30 seconds it is washed off.
After the slide is air-dried it is ready for examination under the
light microscope at not less than .times.800 with oil immersion.
If slides are prepared from tissue impression smears of tumors,
the same procedure is followed but Alexander-Jackson's triple
stain may be applied to duplicate slides in order to differentiate
the non-acid fast forms of Cryptocides from common contaminants.
At times, the Cryptocides organism is not acid-fast in some stages
of its growth.
Usually the organism isolates out in the coccal form which has led
many investigators to believe they are dealing with a
staphylococcus. However, the cocci will be both acid-fast and
non-acid-fast and will vary greatly in shape from the very small
to the large globoidal or sac forms which often stain blue and
appear to be spilling out the red acid-fast cocci much as marbles
out of a bag. In addition, the cocci appear to split
longitudinally into small rods. The cocci, after a period of time,
have small filaments spouting from them which turn into rods that
are red or acid-fast. If the culture is not mutilated by rough
handling, often the large tublar forms can be seen which are
observed by darkfield microscope in fresh blood. These are very
delicate and disrupt easily. The ray formation may also be
apparent but the sheath is extremely diaphanous and is destroyed
often in the staining process. Sometimes the cancer organism can
isolate out primarily as a rod or even as a branching hyphal form.
At other times clublike bodies are seen which are blue in color
and contain the acid-fast bodies within them. It is very important
to study a number of the colonies on the plates and to be sure
that the various transitional forms of the organism can be seen in
one isolated colony. Only then may the organism be grown in
sufficient amount to harvest for the vaccine. All of this work
requires careful examination and experience to be sure of the
growth pattern and morphology of the Cryptocides.
The organisms isolated from the urine cultures have been
classified under various names such as staphylococcus epidermidis
and enterococcus fecalis, in other words, common organisms found
on the skin and in the bowel. However, by careful sterile methods
microorganisms are found to be growing from the urine in great
abundance. Microbiologists are still debating the nature and
classification of these organisms. A recent paper, which appeared
in Transactions of the New York Academy of Sciences, by Dr.
Florence Seibert, claims that these isolates from her material
which yield certain supposedly well-known microorganisms are not
the standard well-recognized types at all but the acid-fast
organisms which we have classified as the Cryptocides.
Microbiologists who examined applicants' urine cultures state that
there are a variety of organisms in applicants' urine vaccines.
What is known is that these organisms occur in large numbers and
are often hemolytic (destructive of red blood cells). Very
possibly urine cultures contain a mixture of whatever microbes
that happen to filter through the kidneys from distant body foci.
Applicants use the mixture in vaccines only as nonspecific immune
booster in chronic disease.
The vaccine can be prepared by the following preferred method. To
10 cc of heparinized freshly-drawn sterile blood add 10 cc sterile
distilled water. Heat over the Bunsen burner to boiling several
times (to break the red cells). Incubate for 10 to 12 days at 37
DEG to 38 DEG C. This gives the growing culture without addition
of media, since blood acts as its own medium without the addition
of anything else. Add 4 percent formalin (formaldehyde) to
inactivate (kill) the P. Cryptocides. The admixture is put through
a microfilter to remove the dead or microorganisms. The live
attenuated microorganisms go through the filters. Then dilute the
filtrate with sterile saline solution until the final solution
contains 1% formalin. This is then tested for sterility and than
can be used as an autogenous vaccine.
The autogenous vaccine can be used to immunize against underlying
chronic diseases or infections, or neoplastic diseased which
produce choroninic gonedotropic hormone and/or biologically
related hormones and sterols which yield a positive indication in
a standard pregnancy test and which can be identified by gas
chromatography and which are caused by Cryptocides.
Materials and Methods
Female mice of strain A/He, 2 months old, were divided into 4
groups of 16 animals each. The average body weight for each group
was 28.1 g. Animals all received 12 intrapertoneal injections, 3
injections per week for 4 weeks (Mondays, Wednesdays and Fridays).
Sterile disposable plastic syringes fitted with 25-gauge needles
were used for each injection.
The mice were housed in plastic shoe box-type cages, 8 per cage,
and fed a standard Teklad mouse diet and water ad libitum. They
were weighed before each injection and then weekly following the
injection period. The injections contain the reddish-brown
crystalline material extracted from broth containing organisims
within the scope of this invention.
Dosages used Dose/Injection
N.C. var. 0.13 mg/0.1 ml
Control 0.13 mg/0.1 ml
#6 1.40 mg/0.1 ml
#5 0.28 mg/0.1 ml
At 20 weeks after the last injection, the mice were killed by
cervical dislocation. Necropsies were performed and the lungs
removed and fixed in Tellyesniczky's fixitive for 24 hours. The
number of tumor nodules on each lung was determined after fixation
by counting with the naked eye.
The animals tolerated the materials extremely well and no adverse
effects were encountered.
J Transl Med. 2006; 4: 14.
Bacteria and cancer: cause, coincidence or
cure? A review
Research has found that certain bacteria are associated with human
cancers. Their role, however, is still unclear. Convincing
evidence links some species to carcinogenesis while others appear
promising in the diagnosis, prevention or treatment of cancers.
The complex relationship between bacteria and humans is
demonstrated by Helicobacter pylori and Salmonella typhi
infections. Research has shown that H. pylori can cause gastric
cancer or MALT lymphoma in some individuals. In contrast, exposure
to H. pylori appears to reduce the risk of esophageal cancer in
others. Salmonella typhi infection has been associated with the
development of gallbladder cancer; however S. typhi is a promising
carrier of therapeutic agents for melanoma, colon and bladder
cancers. Thus bacterial species and their roles in particular
cancers appear to differ among different individuals. Many
species, however, share an important characteristic: highly
site-specific colonization. This critical factor may lead to the
development of non-invasive diagnostic tests, innovative
treatments and cancer vaccines.
An overwhelming body of evidence has determined that relationships
among certain bacteria and cancers exist. The bacterial mechanisms
involved are as yet unclear. These gaps in knowledge make it
impossible to state the exact progression of events by which
specific bacteria may cause, colonize or cure cancer. Therefore,
many questions remain. For example, why do infections that are
wide spread appear to cause cancer in only a minority of
individuals? Do certain infective agents initiate or promote
cancer or does an early undetected cancer facilitate the
acquisition of the infection? Can the exposure to or colonization
of specific bacteria prevent or treat certain cancers? Can the
highly site specific colonization of certain bacteria for a tumor
be clinically useful in the diagnosis of cancer or delivery of a
The scope of this review is broad therefore a wide range of
reports is presented. Recent findings that have found associations
between certain bacterial infections and tumor development will be
discussed as well as genetic factors that may predispose
individuals to "cancer- causing" infections. Mechanisms thought to
be involved with the carcinogenic, diagnostic and preventive or
treatment roles of bacteria are introduced. As the carcinogenic
potential of viral agents and H. pylori has been reviewed
extensively elsewhere, it will not be included here.
Bacteria and carcinogenesis
It is estimated that over 15% of malignancies worldwide can be
attributed to infections or about 1.2 million cases per year.
Pisani et al.  Infections involving viruses, bacteria and
schistosomes have been linked to higher risks of malignancy.
Although viral infections have been strongly associated with
cancers [2,3] bacterial associations are significant. For example,
convincing evidence has linked Helicobacter pylori with both
gastric cancer and mucosa-associated lymphoid tissue (MALT)
lymphoma [4-6], however other species associated with cancers
include: Salmonella typhi and gallbladder cancer [7-10],
Streptococcus bovis and colon cancer [11-14] and Chlamydia
pneumoniae with lung cancer [15-17]. Important mechanisms by which
bacterial agents may induce carcinogenesis include chronic
infection, immune evasion and immune suppression .
It has been shown that several bacteria can cause chronic
infections or produce toxins that disturb the cell cycle resulting
in altered cell growth [15,16,19]. The resulting damage to DNA is
similar to that caused by carcinogenic agents as the genes that
are altered control normal cell division and apoptosis [20,21].
Processes that encourage the loss of cellular control may be tumor
initiators (directly causing mutations) or promoters (facilitating
mutations). Tumorigenesis is initiated when cells are freed from
growth restraints, later promotion results when the immune system
is evaded favoring further mutations and increased loss of cell
control. As the tumor proliferates an increased blood supply is
needed resulting in the organization of blood vessels or
angiogenesis. Subsequent invasion occurs if the tumor breaks down
surrounding tissues. The worst outcome is metastasis which results
when cells break away from the tumor and seed tumors at distant
The immune system is an important line of defense for tumor
formation of malignancies that express unique antigens. Certain
bacterial infections may evade the immune system or stimulate
immune responses that contribute to carcinogenic changes through
the stimulatory and mutagenic effects of cytokines released by
inflammatory cells. These include reactive oxygen species (ROS),
[22,23], interleukin-8 (IL-8) , cyclooxygenase-2 (COX-2),
, reactive oxygen species (ROS) and nitric oxide (NO) .
Chronic stimulation of these substances along with environmental
factors such as smoking, or a susceptible host appears to
contribute significantly to carcinogenesis.
Salmonella typhi and gallbladder cancer
Worldwide annual incidence of gallbladder cancer (GC) is 17
million cases with high incidence rates in certain populations.
The malignancy is usually associated with gallstone disease, late
diagnosis, unsatisfactory treatment, and poor prognosis. The
five-year survival rate is approximately 32 percent for lesions
confined to the gallbladder mucosa and one-year survival rate of
10 percent for more advanced stages . Over 90 percent of
gallbladder carcinomas are adenocarcinoma  involving
gallstones in 78% – 85% of cases .
There are several risk factors for gallbladder cancer. The main
associated risk factors include cholelithiasis (especially
untreated chronic symptomatic gallstones), obesity, reproductive
factors, environmental exposure to certain chemicals, congenital
developmental abnormalities of the pancreatic bile-duct junction
and chronic infections of the gallbladder [26,28]. The interplay
of genetic susceptibility, lifestyle factors and infections in
gallbladder carcinogenesis is still poorly understood ,
however a link has been specifically proposed between chronic
bacterial infections of the gallbladder and Salmonella typhi .
The strongest epidemiological evidence of bacterial oncogenic
potential, aside of Helicobacter pylori, concerns S. typhi.
Infection with this bacterium of typhoid, can lead to chronic
bacterial carriage in the gallbladder . Recent epidemiological
studies have shown that those who become carriers of S. typhi have
8.47 times the increased risk of developing carcinoma of the
gallbladder compared with people who have had acute typhoid and
have cleared the infection . These findings agreed with
earlier investigations by Welton et al.  and Caygill .
A case-control study by Welton et al  compared those who
experienced acute infection with S. typhi to those who
subsequently became chronic carriers following the 1922 typhoid
outbreak in New York. Carriers were six times more likely to die
of hepatobiliary carcinoma than matched controls. Additional
evidence was found in an analysis of the 1964 typhoid outbreak in
Aberdeen . Their findings also suggested a strong association
between chronic carrier status and hepatobiliary carcinoma. These
studies also agreed, people who contracted typhoid but did not
become carriers were not at higher risk of cancer [8,26,30,31].
The highest incidence of gallbladder cancer (GC) in the world is
among populations of the Andean area, North American Indians, and
Mexican Americans. In Europe, the highest rates are found in
Poland, the Czech Republic and Slovakia. The high rates observed
in Latin America are primarily in populations with high levels of
Indian mixture . This evidence supports the notion that
increased susceptibility to gallbladder cancer depends on genetic
factors that predispose people to gallbladder cancer either as
primary factors, or secondarily as promoters by favoring the
development of cholesterol gallstones. The highest mortality rates
are in South America, (3.5–15.5 per 100,000) and among Mexican
Americans . Incidence rates of GC in various ethnic groups in
the USA confirmed the worldwide pattern, as GC was substantially
more frequent among Hispanic than non-Hispanic white women and
men. Interestingly, compared to non-Hispanic whites an excess of
GC was also reported among American Indians in New Mexico, in
agreement with the excess in incidence rates reported for American
Indians and Alaskan Natives . The malignancy is 3 times
higher, however, among women than men in all populations .
Two main pathways to GC exist worldwide. The predominant pathway
involves gallstones and resultant cholecystitis and affects women
to a greater extent than men. The risk of developing gallstones in
response to environmental factors is genetically determined, as
shown by the marked tendency of gallstones to cluster in families
. The other pathway involves an anomalous pancreatobiliary
duct junction (APBDJ), a congenital malformation of the biliary
tract that is more frequent in Japan, Korea, and possibly China,
than in Western countries . In APBDJ, the premature junction
of common bile and pancreatic ducts results in regurgitation of
pancreatic juice into the gallbladder, leading to bile stasis and
inflammation, though generally less severe than that resulting
from gallstones .
Currently the prevention of gallbladder cancer in high risk
populations depends upon the diagnosis of gallstones and removal
of the gallbladder. Indeed, a strong inverse association between
number of cholecystectomies and GC incidence and mortality rates
can be found in many countries. The increase of GC mortality
reported in Chile in the 1980s was related to decreased rates of
cholecystectomies . Increased rates led to the removal of
gallbladders at risk, and a reduction of GC incidence and
mortality in Europe and the United States .
Unfortunately, information about the genetic changes involved in
gallbladder carcinogenesis is limited. Most studies have focused
on gene abnormalities and deletions ("loss of heterozygosity") at
chromosomal regions harboring known or putative tumor suppressor
genes . It appears, however, that TP53 inactivation has an
important and early role in gallbladder carcinoma associated with
gallstones and chronic inflammation. This inactivation would
abrogate the tumor suppressor function of the p53 protein
resulting in impairments in cell cycle control, cellular repair
In contrast, KRAS mutations are frequent and early events in
tumors associated with APBDJ  but detected less often in
gallbladder carcinomas associated with gallstones. KRAS is an
oncogene that encodes a protein that is a member of the small
GTPase family. A mutation in this gene results in an abnormal
protein implicated in several malignancies, including lung
adenocarcinoma, ductal carcinoma of the pancreas and colorectal
carcinoma among others.
Chlamydophila pneumoniae and lung cancer
Lung cancer is the leading cause of cancer death in the United
States and many countries in the Western world. In 2002, the most
recent year for which statistics are available, 90,121 males and
67,509 females died from lung cancer . About 6 out of 10
people with lung cancer die within 1 year of finding out they have
lung cancer. Between 7 and 8 will die within 2 years .
Although patients may experience a partial or complete response to
treatment, most patients relapse and die. Increased dosage of
chemotherapy or length of treatment has not been beneficial .
Chlamydophila (formerly Chlamydia) pneumoniae infection has been
implicated in several chronic lung diseases by serology and direct
antigen detection. Acute lower respiratory tract infection caused
by C. pneumoniae seems often to precede attacks of asthma in both
children and adults but is also involved in some exacerbations of
chronic bronchitis. More importantly it seems to be strongly
associated with chronic obstructive lung disease irrespective of
exacerbation status. Moreover, persistently elevated C. pneumoniae
antibody titers have been observed in sarcoidosis and lung cancer
C. pneumoniae is a Gram-negative bacillus and an intracellular
parasite that causes respiratory infection in more than 50% of
adults. The route of transmission is usually by aerosol and in
most cases these infections are mild. The bacterium is, however,
an important cause of pneumonia, bronchitis, sinusitis, rhinitis
and chronic obstructive pulmonary disease . Respiratory
infections from C. pneumoniae vary in different countries and
populations, being endemic in the United States and epidemic in
Scandinavian countries .
After acute infection the C. pneumoniae intracellular life cycle
is characterized by the development of metabolically inert (and
thus antibiotic resistant) atypical "persistent" inclusions. These
inclusions contain increased quantities of chlamydial heat shock
protein 60, a highly immunogenic protein implicated in the
pathogenesis of chronic chlamydial infections. The resulting
clinical course is acute symptomatic illness followed by chronic
respiratory symptoms. Research also suggests that persistent C.
pneumoniae inflammation correlates with increased risk of lung
cancer [16,17,19]. Prospective and retrospective studies both
report that individuals with elevated IgA antibody titers to this
organism have 50% to 100% increased lung cancer risk .
In a study by Kocazeybek et al.  the relationship between
chronic C. pneumoniae infection and lung carcinoma was examined. A
total of 123 patients who were smokers and diagnosed with lung
carcinoma based on clinical and laboratory (radiological,
cytological) findings were examined. 101 (82.1%) of the cases were
male. 70 had small-cell, 28 squamous-cell and 7 large-cell
carcinomas, while 18 had adenocarcinoma. 123 healthy controls were
matched to the cancer patients by age, gender, duration of smoking
Blood samples (5 ml) were withdrawn at the time of diagnosis (or
enrollment for controls) and 1 month later. Values between IgG
≥512 and IgA ≥40 were set as the criteria for chronic C.
pneumoniae infections. In male patients with lung carcinoma, IgG
antibody titers of ≥512 and IgA antibody titers of ≥40 were found
at a higher rate than in the control group, however, this ratio
was not significant for female patients. These elevations in
antibody titers were found in a total of 62 (50.4 %) cases, 54% of
the male patients and 36% of the female patients. Chronic C.
pneumoniae infections were seen statistically more often in male
patients with carcinoma who were aged 55 years or younger than in
controls (P < 0.001). No difference was reported between male
patients with lung carcinoma over age 55 and controls or in blood
titers between female patients and controls.
The relationship between C. pneumoniae infection and lung
carcinoma was studied by Littman et al.  in a large
prospective case-control study to investigate whether IgA antibody
titers to C. pneumoniae were associated with lung cancer risk. A
total of 508 pairs were enrolled and included both current and
former smokers. Serum was collected at baseline and annually
thereafter. Antibody determinations of each lung cancer subject
and matched control were tested simultaneously in the same
titration series in a blinded fashion. C. pneumoniae titers (IgA
or IgG) ≥16 were considered seropositive, which was consistent
with the cutoff used in other studies. Subjects were matched by
age, gender, and smoking status at baseline. The median age of
cases and controls was 59 years and about half were women. All
subjects were also examined for demographic, lifestyle, dietary,
and racial and ethnic factors. Lung cancer subjects had a heavier
smoking history than controls.
After adjusting for a history of chronic bronchitis or emphysema,
lung cancer subjects were more likely to have IgA titers ≥16
(55.4% vs. 51.3%) and ≥256 (5.1% vs. 2.5%) to C. pneumoniae than
controls. Individuals with antibody tiers IgA ≥16 had 1.2 times
the risk of lung cancer (95% confidence interval, 0.9–1.6)
compared to those with lower titers. Investigators reported a
significant trend (P = 0.007) of increasing odds ratios with
increasing IgA titers primarily due to an odds ratio of 2.8 (95%
confidence interval, 1.1–6.7) associated with titers ≥256.
Elevated IgA was reported with squamous cell carcinomas and to a
lesser extent, for small cell carcinomas and adenocarcinomas.
There was no evidence of a stronger association with elevated IgG
titers however. Subjects with race not classified as White or
Black were more likely to have IgA titers ≥16. No significant
differences in seropositivity were found, however, based on
Streptococcus bovis and colorectal cancer
Colorectal cancer (CRC) is a common malignancy in developed
countries and is the 3rd most common cancer in the United States
. Greater than 80% occur sporadically . The American
Cancer Society estimates that there will be about 104,950 new
cases of colon cancer and 40,340 new cases of rectal cancer in
2005 in the United States. Combined, they will cause about 56,290
deaths. The risk of colon cancer increases after the age of 40 and
rises exponentially from the ages of 50 to 55. In fact, more than
9 out of 10 people found to have colorectal cancer are older than
Survival of CRC is related to the stage of disease at the time of
the initial diagnosis. Between 1985 and 1997, death rates of colon
cancer in the United States declined slightly due to earlier
detection of primary tumors, via stool blood tests, sigmoidoscopy,
colonoscopy, and screening tests for serum carcinoembryonic
antigen concentration (CEA) . The 5-year survival rate for CRC
patients is greater than 90% when tumors are detected at a
localized early stage. After the cancer has spread regionally and
involves adjacent organs or lymph nodes, the rate drops to 40–65%;
survival is less than 10% for patients with distant metastases.
Therefore, there is an urgent need to develop effective treatment
strategies to reduce morbidity and mortality. Surgery is currently
the primary treatment modality for this disease. By the time the
patient presents with recurrent symptoms, however, the disease is
rarely curable by surgery even when combined with other therapies
Several species of bacteria have been linked to chronic infections
of the colon and increased risk of colon cancer including
Escherichia coli  and several streptococci [47,48]. Recent
studies, however, have validated earlier findings of an
association between colon cancer and Streptococcus bovis [11,12].
As early as 1951, McCoy and Mason  suggested a relationship
between colonic carcinoma and the presence of infectious
endocarditis. It was not until 1974  that the association of
Streptococcus bovis and colonic neoplasia was recognized, as
25–80% of patients who presented with a S. bovis bacteremia had a
colorectal tumor. The incidence of S. bovis associated colon
cancer has been determined as 18% to 62% .
S. bovis is a normal inhabitant of the human gastrointestinal
tract that can cause bacteremia, endocarditis, and urinary
infection . Although S. bovis is the 2nd greatest cause of
infectious endocarditis from streptococci , it is frequently
associated with gastrointestinal lesions, especially carcinoma of
the colon [12,51-53]. Notably, the colonic neoplasia may arise
years after the presentation of the condition of bacteremia or
infectious endocarditis .
A retrospective review of forty-five documented cases of S bovis
bacteremia was conducted by Gold et al. . Subjects were
identified by a search of computerized bacteriology records from
one tertiary referral hospital and 1 community hospital located in
the same city. Patient records were reviewed to identify the
presence of colonic neoplasia, the use of gastrointestinal
endoscopy, and the presence of gastrointestinal or extraintestinal
malignancies. Seventeen patients (41% of adult patients) underwent
colonoscopy. Colonic neoplasia was present in 16 patients (39% of
adults). Invasive cancer was present in 13 patients (32% of
adults), 8 of these had malignant lesions arising within the
gastrointestinal tract, 3 affecting the colon and 5 patients had
extraintestinal malignancies. The authors concluded that S. bovis
bacteremia was associated with both colonic neoplasia and
It has been demonstrated that S. bovis or its wall extracted
antigens (WEA) were able to promote carcinogenesis in rats .
In one of these investigations a total of 10 adult rats received
i.p. injections of the carcinogen azoxymethane (AOM) (15 mg/kg
body weight) once per week for 2 weeks. Fifteen days after the
last injection of AOM (week 4) the rats were randomly divided into
three groups. Twice per week during 5 weeks, the rats received, by
gavage either S. bovis (1010 bacteria Group I), WEA (100 μg Group
II) and controls (Group III).
One week after the last gavage (week 10), they found that
administration of either S. bovis or its antigens promoted the
progression of preneoplastic lesions. There were increased
formations of hyperproliferative aberrant colonic crypts, enhanced
expression of proliferation markers and increased production of
IL-8 in the colonic mucosa. Normal rats treated with the bacteria
did not develop hyperplastic colonic crypts, however. The authors
concluded that S. bovis exerts its pathological activity in the
colonic mucosa only when preneoplastic lesions are established.
Under identical experimental conditions Streptococcus gordonii was
substituted for S. bovis. The number of preneoplastic lesions in
the colon of S. gordonii-treated rats was similar to rats treated
with AOM alone (22 ± 2). The authors suggested that S. bovis and
its wall extracted antigens, unlike S. gordonii, act as promoters
of carcinogenesis in a chemically-induced animal model.
In another investigation Biarc et al.  isolated 12 S. bovis
cell-associated proteins (S300) and WEA. Cells of the human
colonic epithelial cell line Caco-2 originally derived from an
adenocarcinoma were grown to confluence and allowed to
differentiate. These cells were stimulated with 200 ul of either
S. bovis WEA (50 μg/ml) or cell-associated proteins S300 (100 μl).
The purified S300 fraction was able to trigger the human cell line
and rat colonic mucosa to release chemokines (human IL-8 or rat
CINC/GRO) and prostaglandin E2 (PgE2). The 12 S. bovis proteins
were highly effective in the promotion of pre-neoplastic lesions
in azoxymethane treated rats. In fact the S300 proteins were able
to induce a 5-fold increase in PGE2 secretion from Caco-2 cells,
as compared with cells stimulated with WEA. The study found that
PGE2 release in the human cells correlated with an over-expression
of cyclooxygease-2 (COX-2).
Evidence has shown that over-expression of COX-2 has a major role
in mucosal inflammation  and is associated with inhibition of
apoptosis  and enhancement of angiogenesis , which favor
cancer initiation and development. It was reported by Biarc et al.
 that S. bovis proteins also promoted cell proliferation by
triggering mitogen-activated protein kinases (MAPKs), which can
increase the incidence of cell transformation, the rate of genetic
mutations and up-regulate COX-2. The investigators concluded that
colonic bacteria such as S. bovis can contribute to cancer
development particularly in chronic infection/inflammation
diseases where bacterial components may interfere with cell
Genetic predisposition to cancer-causing infections
Research has shown that some populations are genetically
predisposed to the infections that are associated with cancer and
indeed have a higher risk of the cancer in question. The exact
mechanisms remain unclear .
E. coli, crohn's disease and colon cancer
Inflammatory bowel disease (IBD) includes both ulcerative colitis
(UC) and Crohn's disease (CD). Both of these disorders have an
increased risk of colorectal cancer (CRC) [38,46,56]. Although
colorectal cancer (CRC) in individuals with IBD only accounts for
1–2% of all cases of CRC in the general population, it is
considered a serious complication of the disease and accounts for
approximately 15% of all deaths in patients with IBD. The
magnitude of the risk has been found to differ, however, in
population-based studies [56-58]. Recent figures suggest that the
risk of colon cancer for people with IBD increases by 0.5–1.0%
yearly, 8–10 years after diagnosis. The magnitude of CRC risk also
increases with early age at IBD diagnosis, longer duration of
symptoms, and extent of disease, with pancolitis having more
severe inflammation and a higher risk of dysplasia-carcinoma
E. coli are found at higher levels in inflammatory bowel disease
(IBD), therefore, studies have examined the mechanisms that may
explain this phenomenon. A cell culture study by Martin et al 
attempted to quantify and characterize mucosa-associated and
intramucosal bacteria, particularly E. coli, in these inflammatory
conditions. Their hypothesis was that the disease-associated
alterations in mucosal glycosylation found in inflammatory bowel
disease and colon cancer might predispose to altered recruitment
of bacteria to the mucosa.
Mucosa-associated bacteria were isolated from biopsy samples of
Crohn's disease, (n = 14); ulcerative colitis, (n = 21);
noninflamed controls, (n = 24) and at surgical resection of colon
cancer, (n = 21). Results found that mucosa-associated and
intramucosal bacteria were cultured more commonly in Crohn's
disease (79%, P = 0.03; and 71%, P < 0.01, respectively), and
colon cancers (71% and 57%) than in noninflamed controls (42% and
29%) but not ulcerative colitis (38% and 48%). Mucosa-associated
E. coli, which accounted for 53% of isolates, were more common in
Crohn's disease (6/14; 43%) than in noninflamed controls (4/24,
17%), and intramucosal E. coli more common in Crohn's disease
(29%; controls, 9%).
E. coli expressed hemagglutinins in 39% of Crohn's cases and 38%
of cancers but only 4% of controls, and this correlated (P = 0.01)
with adherence to embryonic intestinal cells (I407) and colon
adenocarcinoma cells (HT29). Although close apposition of E. coli
resulted in release of pro-inflammatory cytokines, cellular
invasion by bacteria was not essential to this process .
Aspinell  suggested that the bacterial adherence found by
Martin et al.  might result from activation of virulence genes
following contact of the organisms with the inflamed mucosal
cells. Martin et al.  found, however, that the mucosal
isolates expressed of none of the known virulence genes, other
than adherence genes. Martin and co-workers concluded that their
findings supported a central role for mucosally adherent bacteria
in the pathogenesis of Crohn's disease. They postulated that
similar, lower grade, inflammatory changes could contribute to the
risk of sporadic cancer development .
The authors stated however that it was certainly possible that the
presence of the bacteria in the sub-mucus niche in human Crohn's
disease and colon cancer could have been encouraged by
disease-associated changes  in the mucosa. If true, their
findings would result from colonization coincidental to the
disease-associated alterations in mucosal glycosylation found in
inflammatory bowel disease and colon cancer.
A study conducted by Masseret et al. examined the E. coli
strains isolated from patients with Crohn's disease (CD) with
chronic ileal lesions (n = 14), early endoscopic recurrent lesions
(n = 20), without endoscopic recurrence (n = 7), and controls (n =
21). Genetically linked E coli strains were isolated significantly
more frequently from patients with chronic and recurrent CD (24/33
patients) than from controls (9/21) (p < 0.05). Most patients
operated on for chronic ileal lesions (78.5%) harbored E coli
strains belonging to the same cluster (p < 0.002 v controls).
The prevalence of patients with early recurrent lesions harboring
E coli strains belonging to this cluster was high but not
significant. 21 of 26 strains isolated from patients with active
CD demonstrated adherent ability to differentiated Caco-2 cells,
indicating that most of the genetically related strains shared a
common virulence trait. Comparison of E coli strains recovered
from ulcerated and healthy mucosa of patients operated on for CD
demonstrated in each patient that a single strain colonized the
intestinal mucosa. The authors suggested that although a single E
coli isolate was not found in Crohn's ileal mucosa, some genotypes
were more likely than others to be associated with chronic or
early recurrent ileal lesions.
S. typhi and susceptible populations
As previously stated, certain populations have an increased risk
of gallbladder cancer (GC), however certain individuals may be
predisposed to S. typhi infection which appears to increase the
risk of GC. In an investigation by deJong et al. , three
unrelated individuals with severe, idiopathic mycobacterial and
Salmonella infections were found to lack IL-12Rβ1 chain
expression. Interleukin-12 (IL-12) is a cytokine that promotes
cell-mediated immunity to intracellular pathogens, such as S.
typhi, by inducing type 1 helper T cell (TH1) responses and
interferon-γ (IFN-γ) production. IL-12 binds to the high-affinity
β1/β2 heterodimeric IL-12 receptor (IL-12R) complexes on T cell
and natural killer cells. The cells of these patients were
deficient in IL-12R signaling and IFN-γ production and their
remaining T cell responses were independent of endogenous IL-12.
IL-12Rβ1 sequence analysis revealed genetic mutations that
resulted in premature stop codons in the extracellular domain. The
genetic absence of IL12-Rβ1 expression represented an immune
deficiency in these 3 patients. Interestingly, these patients did
not develop any abnormal infections with other viral, bacterial,
or fungal pathogens. The defect in IFN-production and extreme
susceptibility to mycobacterial and Salmonella infections in these
patients appeared to be a direct result of their lack of IL-12R
expression and signaling. The authors concluded that selective
susceptibility to mycobacterial and Salmonella infections,
however, suggested that the type-1 cytokine pathway was essential
for controlling resistance to the intracellular pathogens and that
no redundant protective immune mechanism could compensate for this
Respiratory conditions and increased susceptibility to lung
75–90% of people who develop lung cancer are smokers, however,
only a small proportion of smokers develop lung cancer .
Hence, epidemiological studies such as that of Littman et al 
and Kocazeybek et al.  have been conducted to more closely
identify risk factors. Identifying genetic factors that increase a
smoker's risk of developing lung cancer may help scientists to
better understand the etiology of lung cancer and more effectively
target high-risk groups for screening. Additionally, genetic
factors have been identified that appear to predict the prognosis
of certain lung cancer patients . For example, mutations
affecting the epidermal growth factor receptor (EGFR) were
significantly associated with specific genetic alterations.
Supervised clustering analysis based on EGFR gene mutations
elucidated a subgroup including all EGFR gene mutated tumors,
which showed significantly shorter disease-free survival
To analyze the genetic alterations of primary lung adenocarcinoma
in a high-throughput way, Shibata et al.  used laser-capture
micro-dissection of cancer cells and array comparative genomic
hybridization focusing on 800 chromosomal loci containing
cancer-related genes. They identified a large number of
chromosomal numerical alterations, including frequent
amplifications. Three subgroups of lung adenocarcinoma were
characterized by distinct genetic alterations and were associated
with smoking history and gender. The authors concluded that
multiple carcinogenic pathways exist; certain abnormalities appear
related to gender and smoking while others may impact survival
Bacterial strategies: cell cycle control and toxic warfare
Bacterial toxins can kill cells or at reduced levels alter
cellular processes that control proliferation, apoptosis and
differentiation. These alterations are associated with
carcinogenesis and may either stimulate cellular aberrations or
inhibit normal cell controls. Cell-cycle inhibitors, such as
cytolethal distending toxins (CDTs) and the cycle inhibiting
factor (Cif), block mitosis and are thought to compromise the
immune system by inhibiting clonal expansion of lymphocytes. In
contrast, cell-cycle stimulators such as the cytotoxic necrotizing
factor (CNF) promote cellular proliferation and interfere with
cell differentiation .
Bacterial toxins that subvert the host eukaryotic cell cycle have
been classified as cyclomodulins. For example, CNF is a cell-cycle
stimulator released by certain bacteria, such as E. coli. CNF
triggers G1 – S transition and induces DNA replication. The number
of cells does not increase, however. The cells become
multinucleated instead, perhaps by the toxin's ability to inhibit
cell differentiation and apoptosis [63,64].
Conversely the cytolethal distending toxin (CDT), as previously
mentioned, is a cell-cycle inhibitor used by several species of
Gram-negative bacteria, including Campylobacter jejuni and S.
typhi. The CdtB unit of CDT is a DNAse that creates
double-stranded DNA breaks causing cell cycle arrest, usually at
the G2 checkpoint . Cif is a cell cycle inhibitor found in
enteropathogenic (EPEC) and enterohaemorrhagic (EHEC) E. coli.
EPEC and EHEC deliver this novel toxin by injecting it into the
infected epithelial cells. Cif arrests the cells at the G2/M phase
causing unique alterations in the host cell that result in
attachment of the cytoskeleton to the host cell membrane. This
anchoring of the cytoskeleton inhibits mitosis, causing cellular
and nuclear enlargement. Although DNA synthesis is initiated it
does not lead to nuclear division. Endoreduplicaton occurs
resulting in cellular DNA content of 8–16n [20,66].
In a cell culture study, Haghjoo and Galán  found that S.
typhi produced a unique cdtB-dependent CDT that required bacterial
internalization into host cells. When Cos-2 cells were transfected
with S. typhi the effects of the cdtB subunit were severe
fragmentation of chromatin characteristic of the CdtB subunit of
CDT expressed by other species. The authors proposed that S. typhi
subsequent to internalization deviated from the usual endocytic
pathway that leads to lysosomes, reaching an unusual
membrane-bound compartment where it can survive and replicate. It
is possible that this unique CDT may be involved in some aspects
of the ability of S. typhi to cause long, persistent infections in
humans, because, at least in other bacteria, this toxin has been
shown to possess immunomodulatory activities.
Toxins are not the only strategy for evading the host's immune
system, however. An early study by Kilian et al.  reported
that some strains of Capnocytophaga ochracea, an oral pathogen,
are capable of hydrolytically degrading immunoglobulin A subclass
1 found in the oral cavity. This property may enhance colonization
and invasion of oral lesions which characterize many bacteremias
due to Capnocytophaga species. . Shurin et al.  obtained
evidence that Capnocytophaga species inhibit polymorphonuclear
leukocyte migration; a means by which these species may evade
The immune system may also be evaded by the protection offered by
bacterial biofilms. An example of this phenomenon is provided by
uropathic E. coli species whose biofilms protect it from the
immune system and making it difficult to treat these infections
effectively by antibiotics. This has been demonstrated in bladder
infections where the same species is recovered after repeated
flare-ups thought to have been cleared by antibiotic therapy,
suggesting a subclinical infection that has become chronic .
Bacterial site-specific colonization
Bacterial adherence is thought to be the first important step in
colonization. It is now recognized that bacteria bind to and
colonize host cells in a highly selective manner via a "lock- and
key" mechanism. This selectivity of bacterial adhesion plays an
important role in many infectious processes, and an understanding
of the mechanisms involved could provide molecular explanations
for the innate resistance or susceptibility of hosts and tissues
to many infectious agents.
Regulators of complement activation (RCA proteins) prevent the
destructive consequences of inappropriate immune activation.
Decay-accelerating factor (CD55) is a member of the RCA protein
family that protects host cells from complement damage and
regulates the classical, alternative and lectin pathways that
converge to target cells for destruction in all 3 pathways of the
innate immune system . CD55 is expressed on all serum-exposed
cells. Perhaps due to its ubiquitous expression, it is thought
that bacterial pathogens, including uropathogenic Escherichia
coli, use CD55 as a receptor prior to infection. Williams et al.
 suggested that pathogens have evolved to exploit the cellular
roles of this molecule thereby gaining immunological advantage
The influence on E. coli binding of the two known single amino
acid polymorphisms within short consensus repeat (SCR) domains of
CD55 was examined by Pham et al.  and Nowicki et al . The
bacterial strains sensitive to a change in SCR3 were found to be
insensitive to changes in SCR4 and vice versa, suggesting that
multiple, independent binding sites of CD55 were used by different
bacterial strains. Evidence from those investigations suggested
that E. coli strains sensitive to changes in one binding domain
were not affected by changes in other domains. Furthermore, the
use of CD55 as a receptor by a variety of uropathic E. coli was
found to correlate with symptomatic infections [71,72]. Evidence
from those investigations indicated the extraordinary degree of
site-specific colonization of these closely related strains.
Bacteria associated with a coincidental or diagnostic role
Each year nearly 30,000 Americans are diagnosed with oral cancer
[73,74]. Over 90% of these malignancies are oral squamous cell
carcinoma (OSCC). Despite advances in surgery, radiation and
chemotherapy, the five-year survival rate is 54%, one of the
lowest of the major cancer sites and this rate has not improved
significantly in recent decades [38,75,76]. The disease kills one
person every hour – more people than cervical cancer, Hodgkin's
disease, or malignant melanoma . Notably, incidence in young
adults (<40 years) is increasing in the U.S. [8,10] and
worldwide [9,77]. The World Health Organization predicts a
continuing worldwide increase in oral cancer over the next several
Early detection followed by appropriate treatment, increases cure
rates to about 80%, and greatly improves the quality of life by
minimizing extensive, debilitating treatments . Oral cancer is
asymptomatic in its early stages, however, and in spite of the
accessibility of the oral cavity to direct examination, these
malignancies are often not detected until a late stage [79-81].
Oral cancer is unusual in that it carries a high risk of second
primary tumors. Patients who survive a first cancer of the oral
cavity have up to a 20-fold increased risk of developing a second
primary oral cancer. The heightened risk can last 5–10 years,
sometimes longer .
In response to the difficulties in effectively treating oral
cancer, research studies are focusing on prevention and early
diagnostics. Some of these studies have found that OSCC lesions
are colonized by an altered microbiota [83,84]. Other
investigations have found bacterial DNA or live organisms within
oral cancer tissues [85,86]. The true nature of the relationships
between oral bacteria and oral or esophageal cancers is, however,
PCR techniques have been used to seek the DNA of bacterial species
in head and neck cancer tissues. Sasaki et al.  found S.
anginosus DNA sequences in tissue samples from 127 cancer
patients. Tissues examined included esophageal cancer, gastric
cancer tissues, and dysplasia of the esophagus from esophageal
cancer patients. No S. anginosus DNA was found in noncancerous
esophagus or stomach samples. However, the degree of S. anginosus
infection in biopsied tissues was much more obvious in the
dysplastic and cancerous sections than in the noncancerous
portions of the esophagus suggesting that S. anginosus infection
occurred at an early stage of esophageal cancer. The authors
suggested that S. anginosus could play a significant role in the
carcinogenic process of most cases of esophageal cancer and some
cases of gastric cancer by causing inflammation.
Morita et al.  found that 8 of 18 (44%) samples from the
esophagus contained a detectable level of S. anginosus DNA, but
only 5 of 38 (13%) of oral cancer had detectable DNA levels of
this organism. The quantity of S. anginosus DNA in the esophageal
cancer tissues was significantly higher than in oral cancer. The
maximum amount of S. anginosus DNA was approximately 10 times
higher in esophageal than in oral cancer tissues. In addition,
none of the 5 different oral cancer sites (floor of mouth,
maxillary or mandibular gingiva, buccal mucosa, and tongue) showed
significant signs of S. anginosus infection. Most non-cancerous
tissues of the esophagus and tongue showed an undetectable level
of S. anginosus. The authors concluded that S. anginosus is
associated with esophageal cancer, but is not closely related with
In a previous study by Mager et al.  it was determined that
the salivary microbiota was similar to that of the oral soft
tissues. Therefore, the investigators examined whether the
salivary counts of 40 common oral bacteria in subjects with an
oral squamous cell carcinoma (OSCC) lesion would differ from those
found in cancer-free (OSCC-free) controls . Unstimulated
saliva samples were collected from 229 OSCC-free and 45 OSCC
subjects and evaluated for their content of 40 common oral
bacteria using checkerboard DNA-DNA hybridization.
DNA counts per ml saliva were determined for each species,
averaged across subjects in the 2 subject groups, and the
significance of differences between groups determined using the
Mann-Whitney test and adjusted for multiple comparisons. The
diagnostic sensitivity and specificity in detection of OSCC by
levels of salivary organisms were computed and comparisons made
separately between a non-matched group of 45 OSCC subjects and 229
controls and a group of 45 OSCC subjects and 45 controls matched
by age, gender and smoking history.
Counts of 3 of the 40 species tested, Capnocytophaga gingivalis,
Prevotella melaninogenica and Streptococcus mitis, were elevated
in the saliva of individuals with OSCC (p < 0.001). When tested
as diagnostic markers the 3 species were found to predict 80% of
cancer cases (sensitivity) while excluding 83% of controls
(specificity) in the non-matched group. Diagnostic sensitivity and
specificity in the matched group were 80% and 82% respectively.
These findings suggest that high salivary counts of C. gingivalis,
P. melaninogenica and S. mitis could be diagnostic indicators of
The reasons for the differences in colonization patterns of
specific bacterial species at different host locations are only
partially understood. These reasons include differences in
nutrient availability, competition among species for binding
sites, inter-species antagonisms or cooperations, and the
differences in receptors present on different tissues that permit
binding by specific adhesins possessed by different species. Other
factors that may partly explain the unfavorable microbial shifts
observed in oral carcinoma surface biofilms are a compromised host
response or the irregularity of the lesion surface providing
The most intensely studied of these possibilities has been the
specificity in adhesion of different bacterial species to
receptors on oral soft tissues. Many studies have focused on
fimbriae-mediated adhesion and adhesins in the adherence of
different oral species to oral epithelial cells [88-91]. As a
universal trait of cancer cells is alterations in cell surface
receptors, studies have examined the colonization of healthy and
cancerous epithelia [83,85-87,92].
A study by Neeser et al.  examined the binding of a common
oral bacterial species, Streptococcus sanguis OMZ 9 to healthy and
cancerous buccal cell lines. Results showed that S. sanguis bound
to exfoliated human buccal epithelial cells in a sialic
acid-sensitive manner. The desialylation of such cells invariably
abolished adhesion of S. sanguis to the epithelial cell surface.
The resialylation of desialylated HBEC with CMP-sialic acid and
Galß1,3GalNAc α2,3-sialyltransferase specific for O-glycans
restores the receptor function for S. sanguis OMZ 9, whereas a
similar cell resialylation with the Galß1,4GlcNAc
α2,6-sialyltmnsferase specific for N-glycans is without effect.
These findings suggested that a 23 kDa cell surface glycoprotein
bearing a carbohydrate sequence, NeuNAc alpha 2-3Gal beta
1-3GalNAc O-linked sugar chains, is recognized by S. sanguis on
exfoliated human buccal epithelial cells. In similar experiments
carried out with a buccal carcinoma cell line termed SqCC/Y1, S.
sanguis did not attach in great numbers to cultured tumor cells.
These cells were shown to not express the membrane glycoprotein
bearing alpha 2,3-sialylated O-linked carbohydrate chains.
Aberrations in the cell surface carbohydrate structures have now
been established as a universal characteristic of malignant
transformation of cells, and cancer has been referred to as a
molecular disease of the cell membrane glycoconjugates [93,94].
Thus, changes in the tumor cell surface structure could alter the
adhesion of different species of oral bacteria. Notably, even
species within the same genera, such as streptococci, have been
found to differ in their colonization of healthy and cancerous
oral tissues [83,87].
Bacteria and the prevention or treatment of cancer
Evidence is mounting that certain species of bacteria or their
toxins may indeed have a protective or curative role in some
cancers. Factors that would suggest a protective role of a
bacterial species include: (1) colonization lowers the risk of a
certain cancer; (2) elimination or absence of colonization raises
the risk, or (3) introduction of the bacteria or its toxins cures
or causes remission of the cancer.
Tumors and coley's toxins
Spontaneous tumor regression has followed severe bacterial,
fungal, viral and protozoal infections. For hundreds of years this
phenomenon inspired the development of the earliest cancer
therapies. Reports of spontaneous remissions of advanced cancers
infections can be found in the late nineteenth and early twentieth
centuries. Many of these unexplained cures followed bacterial
infections accompanied by high fevers.
An American surgeon, Dr. William Coley began the first
well-documented use of bacteria and their toxins to treat end
stage cancers. Coley first used live Streptococcus pyogenes
cultures. Problems with the predictability of patient responses
caused him to develop a safer vaccine in the late 1800's composed
of two killed bacterial species, S. pyogenes and Serratia
marcescens. In this way he could simulate an infection with the
accompanying fever without the risk of an actual infection
Coley's vaccine was widely used to successfully treat sarcomas,
carcinomas, lymphomas, melanomas and myelomas. Complete, prolonged
regression of advanced malignancy was documented in many cases.
The combined reports of Coley and others estimated the 5-year
survival rate at 80% in malignancies for which no treatment
existed. Even in patients considered in the terminal stages of
cancer some remarkable recoveries were reported with the patient
often outliving the cancer .
Coley considered 4 points critical to success: (1) initiation of a
naturally occurring infection with fever, (2) avoidance of immune
tolerance by gradually increasing the dosage, (3) directly
injecting the vaccine into the tumor when accessible, and (4) a
minimum of 6 months of injections to avoid recurrences. Today
little credence is given to the febrile response in fighting
A retrospective study was conducted in 1999 to compare the 10 year
survival rate of patients treated by Coley's vaccine with modern
conventional therapies. Richardson et al.  tried to match 128
of Coley's cases with 1,675 controls from the Surveillance
Epidemiology End Result (SEER) cancer registry. The 2 populations
were matched by age, gender, ethnicity, stage and radiation
treatment status. Limitations included sample size and staging of
patients receiving Coley's vaccine. The authors concluded that
"Given the tremendous advances in surgical techniques and medicine
in general, any cohort of modern patients should be expected to
fare better than patients treated 50 or more years ago. Yet no
such statistical advantage for the modern group was observed in
this study." These findings were supported by case reports of
spontaneous remissions or significant benefits when accidental
infections occurred [98-100].
What role may a febrile response play in the remission of a tumor?
Hobohm  offers the following hypothesis. Fever causes a
cascade of events of inflammatory factors which activate resting
dendritic cells (DC) that lead to the activation of T-cells.
Cancer-cell specific T-cells usually remain in a state of anergy,
most likely due to the absence of danger signals that usually
accompany tissue destruction and inflammation upon acute infection
. A feverish bacterial infection may have a 3-fold beneficial
effect. First, many infectious agents release endotoxins, like
LPS, induce inflammatory cytokines and stimulate DC. Second, both
thymocyte proliferation and generation of allo-specific CTL are
increased with temperature in vitro . Third, the vasculature
of a tumor is more fragile than that of normal tissues and
therefore more prone to destruction by the immune response. An
infection causing hemorrhagic necrosis could trigger febrile
collapse of the tumor vasculature [104,105]. Interestingly, the
affinity of certain streptococci for binding to fibrinogen and
fibrin may account for the 'homing' of bacterial enzymes to tumors
as these cells are abundant in such proteins .
The mechanism by which infection cures cancer has been
investigated. It has been suggested by Zacharski and Sukhatme 
that the tumor regression observed by Coley and others is due to
the activation of plasminogen. For example when the streptococcal
spreading factor known as streptokinase (SK) combines with host
plasminogen, plasmin is released. Plasmin triggers protease
cascades that degrade plasma and extracellular matrix proteins.
These mechanisms of degradation are toxic to tumor cells, disrupt
the tumor extracellular matrix, alter tumor growth and inhibit
metastasis . The notion that plasminogen activators like SK
might result in the remissions reported by Coley is appealing as
they appear to spare healthy cells while attacking tumors.
Zacharski et al.  hypothesized that although the potent
enzymes produced by plasminogen activation may have a direct
effect on cancer cells it was more likely they disrupted the
cell-extracellular matrix of the tumor.
Investigators report that the traditional best treatment options
for some candidate tumor types, such as advanced soft tissue
sarcomas, breast cancer and melanoma, have not improved patient
outcome substantially since Coley's day [96,108,109]. Currently,
biologic response modifier therapies have moved beyond the
nonspecific immunotherapy of Coley's era and laid the foundation
for today's approaches. Zarcharski and Sukhatme  suggest that
the early success of Coley's toxins are leading to therapies that
engage the host's immune system against an individual's tumor
offering new hope for cancer patients.
Autologous tumor cell vaccine therapy is an example of this new
approach to cancer treatment. These vaccines differ markedly from
conventional cytotoxic drug therapy that affect both normal and
tumor cells. Tumor vaccines stimulate an individual's
cell-mediated immune response by targeting the patient's tumor
antigens. While efficacy of standard chemotherapy relates to the
dose of the drug, the efficacy of a tumor vaccine is more complex,
involving host-vaccine interactions . These include: (1)
immunogenicity of the vaccine regarding tumor-associated antigens
as opposed to self; (2) the host's immune response in terms of
immune recognition and effector mechanisms; and (3) the
development of host systemic cell-mediated immunity, including
long-term immunologic memory, (3–5 years). Therefore, the potency
of the vaccine is not determined by immunogenicity alone but by
its ability to induce the host anti-tumor response .
Bacillus calmette-guérin and autologous tumor cell vaccine vs.
Certain tumor antigens are, however, normally weak immunogens.
Therefore the use of adjuvants and the intradermal route of
injection have, in some cases, produced an optimum antigenic
vaccine. These adjuvant vaccines have induced effective host
recognition of tumor-associated antigens and improved patient
survival. For example, preliminary evidence by Hoover et al 
suggested that active specific immunotherapy (ASI) of colon cancer
using autologous tumor cell vaccines had potential in improving
recurrence-free interval and survival. ASI assumes there are
distinct tumor antigens on an individual's cancer cells that are
either absent or in lower concentration on normal cells. The
vaccine attempted to stimulate host's immune defenses against
tumor-associated antigens by enhancing the immunogenicity of the
patient's own tumor cells with an immunomodulating adjuvant, such
as Bacillus Calmette-Guérin (BCG).
In a study by Hoover et al.  80 eligible subjects with colon
(47) or rectal (33) cancer were enrolled into a prospectively
randomized, controlled clinical trial of active specific
immunotherapy (ASI). An autologous tumor cell-Bacillus Calmette-
Guerin (BCG) vaccine was used to determine whether ASI could
improve disease- free status and survival. Eligible subjects had
colon or rectal cancers extending through the bowel wall or had
positive lymph nodes providing adequate cells from the primary
tumor. Wide surgical removal of all tumors was performed with
histologically proven clear margins and removal of involved lymph
nodes. Prior to randomization individuals were screened for
metastatic disease. Colon cancer and rectal cancer subjects were
in separate but parallel studies and randomized into groups
treated by resection alone or resection plus ASI. 3–4 weeks
following surgery, both controls and treatment subjects were skin
tested for immune competence and sensitivity to tuberculin
purified protein derivative (PPD). Vaccines were begun in the ASI
treatment group 4–5 weeks following surgery to allow for adequate
immune recovery from surgery and anesthesia. A total of 24 colon
and 17 rectal subjects composed the treatment group. With a median
follow-up of 93 months, there was a significant improvement in
survival (two-sided P = .02; hazards ratio, 3.97) and disease-free
survival (two-sided P = .039; hazards ratio, 2.67) in all eligible
colon cancer patients who received ASI. With a median follow-up of
58 months, no benefits were seen in patients with rectal cancer
who received ASI. The authors concluded that the study suggested
that ASI may be beneficial to patients with colon cancer.
In 2005, Uyl-de Groot et al.  conducted a multicenter,
randomized controlled phase III clinical trial with Stage II and
III colon cancer patients using ASI. Autologous tumor cells were
used with the immunomodulating adjuvant Bacillus Calmette-Guérin
(BCG) in a vaccine (OncoVAX®). Patients were randomized to receive
either OncoVAX® or no therapy after surgical resection of the
primary tumor. The vaccine was processed within 48 h after surgery
in order to have viable, metabolically active, autologous tumor
Analysis of prognostic benefit with a 5.8 year median follow-up,
showed that the beneficial effects of OncoVAX® were statistically
significant at all endpoints including recurrence-free interval,
overall survival, and recurrence-free survival in Stage II colon
cancer patients. Surgery alone cures 65% of Stage II colon cancer
patients. For the remaining patients, OncoVAX® in an adjuvant
setting significantly prolongs recurrence-free interval and
significantly improves 5-year overall survival and recurrence-free
survival. Unfortunately, no statistically significant prognostic
benefits were achieved in Stage III patients .
Immunization with bacillus calmette-guérin vs. lung cancer
Grant et al  hypothesized that optimal chemotherapy with or
without radiation followed by active immunization could eliminate
microscopic residual disease and prolong survival. Immunization
with GD3, a ganglioside expressed on the surface of most small
cell lung cancers (SCLC) had not evoked a strong immune response.
Therefore BEC2, a large xenogenic protein which mimics GD3, was
judged to be a good immunogenic candidate. This approach had
proven successful in extending the lives of melanoma patients
Chapman et al.  conducted a phase II trial comparing 5 dose
levels of BEC2. The study population consisted of 15 patients with
small cell lung cancer (SCLC). All subjects had completed standard
therapy and had achieved a partial or complete response. Patients
received a series of five intradermal immunizations consisting of
2.5 mg of BEC2 plus BCG over a 10-week period. Blood was collected
for serological analysis, and outcome was monitored. All patients
developed anti-BEC2 antibodies, despite having received
chemotherapy with or without thoracic radiation. Anti-GD3
antibodies were detected in five patients, including those with
the longest relapse-free survival. The median relapse-free
survival for patients with extensive stage disease was 10.6
months. In patients with limited stage disease a median
relapse-free survival had not been reached with a follow-up of
>47 months and only one of the 7 patients in this group
relapsed. The authors reported that immunization of SCLC patients
using BEC2 plus BCG after standard therapy could induce anti-GD3
antibodies and was safe. The survival and relapse-free survival in
this group of patients was substantially better than those
observed in similar patients receiving standard therapy.
A Phase III trial was conducted to evaluate BEC2 plus BCG as
adjuvant therapy for limited small-cell carcinoma after
chemotherapy and irradiation . A total of 515 subjects were
randomly assigned. Unfortunately, in this trial there was no
improvement in survival, progression-free survival, or quality of
life in subjects that were vaccinated. A trend toward prolonged
survival was observed in the one third of subjects who developed a
humoral response (p = 0.085), however.
The effectiveness of vaccines for several cancers was examined in
a series of investigations. Xiang et al.  tested vaccines for
human melanoma using the mutant S. typhi strain SL7207 as a DNA
carrier. Tolerance against self-antigens was broken by genetically
fusing ubiquitin with MHC I derivatives. Another approach coupled
tumor-specific antibodies to functional IL-2. This combination in
addition to oral vaccination with plasmid-encoded tumor antigens
significantly enhanced protection against carcinoma of the colon
, carcinoma of the lung  and melanoma . Unstable
cancer cells provided a challenge, however. Interestingly, this
was overcome by targeting stable, proliferating endothelial cells
of the tumor vasculature. This novel approach effectively
inhibited angiogenesis .
Helicobacter pylori and esophageal adenocarcinoma
In industrialized countries the incidence of H pylori has been
steadily decreasing . The incidence of esophageal cancer
(EA), however, is increasing . Surprisingly, there is
evidence that these two trends may be related. Several studies
have determined that virulent strains of H pylori are found less
commonly among patients with Barrett's esophagus and EA when
compared with controls [122-124]. This led to studies that found
positive associations among the increased incidence of obesity,
GORD, Barrett's esophagus and EA .
Recently, a nested case-control study was conducted by de Martel
et al  to assess the association between H. pylori infection
and the risk of development of EA. Of a total of 128,992 members
of an integrated health care system who had participated in a
multiphasic health checkup (MHC) during 1964–1969, 52 patients
developed EA during follow-up. Three randomly chosen control
subjects from the MHC cohort were matched to each cancer subject,
on the basis of age, gender, race, date and site of the MHC. Data
on cigarette smoking, alcohol consumption, body mass index (BMI),
and education level were obtained. Serum samples collected at the
MHC were tested for IgG antibodies to H. pylori and to the H.
pylori CagA antigen associated with H. pylori virulence.
Subjects with H. pylori infections were less likely than
uninfected subjects to develop EA odds ratio (OR, 0.37) 95%
confidence interval (CI, 0.16–0.88). This significant association
was restricted to cancer subjects and control subjects <50
years old (OR, 0.20) (95% CI, 0.06–0.68). Interestingly, in
patients with H. pylori infections, the OR for EA in those who
tested positive for IgG antibodies to the CagA protein was similar
to that for those who tested negative for it. BMI ≥25 and
cigarette smoking, however, were strong independent risk factors
for EA. The authors found, however, that the absence of H. pylori
infection, independent of cigarette smoking and BMI, was
associated with an increase in the risk of development of EA
An epidemiological study in Sweden sought to determine whether BMI
was associated with esophageal malignancies compared to gastric
adenocarcinoma and controls. In a nationwide, population-based
case-control study by Lagergren et al , between 1995 through
1997, a total of 189 patients with adenocarcinoma of the esophagus
and 262 patients with adenocarcinoma of the gastric cardia were
enrolled. These patients were compared with 167 patients with
incident esophageal squamous cell carcinoma and 820 healthy
Odds ratios were determined from BMI and cancer case-control
status and ratios estimated the relative risk for the two
adenocarcinomas studied. Calculations used multivariate logistic
regression with adjustment for potential confounding factors. The
adjusted odds ratio was 7.6 (95% CI, 3.8 to 15.2) among persons in
the highest BMI quartile compared with persons in the lowest.
Obese persons (persons with a BMI>30 kg/m2) had an odds ratio
of 16.2 (CI, 6.3 to 41.4) compared with the leanest persons
(persons with a BMI<22 kg/m2). The odds ratio for patients with
cardia adenocarcinoma was 2.3 (CI, 1.5 to 3.6) in those in the
highest BMI quartile compared with those in the lowest BMI
quartile and 4.3 (CI, 2.1 to 8.7) among obese persons. Although a
strong dose-dependent relation existed between BMI and esophageal
adenocarcinoma, esophageal squamous-cell carcinoma was not
associated with BMI. A modest but significant increase in
intragastric acidity was also observed following the cure of H
pylori infection which the authors postulated could contribute to
gastroesophageal reflux disease (GORD).
The incidence of EA has increased rapidly over the last 30 years.
During this period, the prevalence of Helicobacter pylori has
decreased. Trends of increasing esophageal adenocarcinoma can be
linked causally to increasing GORD which can be linked to an
increasingly obese population. There appeared to be no plausible
biological mechanism of association between H pylori, obesity, and
GORD until studies of ghrelin, however.
Ghrelin was the first circulating hormone demonstrated to
stimulate food intake in man. This peptide is produced in the
stomach and regulates appetite, food intake, and body composition.
The effects of ghrelin were examined in H pylori positive
asymptomatic subjects by several investigators [128-130]. In a
randomized double-blind cross-over study, by Wren et al. ,
ghrelin was shown to acutely enhance appetite and increase food
intake in 9 healthy human subjects. There was a clear-cut increase
in calories consumed by every individual from a free-choice buffet
(mean increase 28 +/- 3.9%, p < 0.001) during ghrelin versus
saline infusions. Furthermore, visual analogue scores for appetite
were greater during ghrelin compared to saline infusion. Ghrelin
had no effect on gastric emptying, however. The authors concluded
that endogenous ghrelin was a potentially important new regulator
of the complex systems controlling food intake and body weight.
Evidence is accumulating that ghrelin may explain the relative
rarity of H. pylori among patients with Barrett's esophagus and
EA. Findings from these studies and others support the notion that
H. pylori may have a "protective" effect against EA [122,124].
Studies have found that curing H pylori infection increased plasma
ghrelin in healthy asymptomatic subjects which may lead to
increased appetite, weight gain and contribute to the increasing
obesity seen in Western populations where the prevalence of H
pylori is low. This evidence supports the notion that decreasing
incidence of H pylori infection may lead to increased levels of
plasma ghrelin and that this hormone appears to be a factor in
increasing obesity which elevates the risk of GORD which is
positively associated with Barrett's esophagus and increased risk
of esophageal adenocarcinoma. It appears that the absence of H.
pylori infection may be one of several factors that leads to the
increased incidence in EA effect observed in Western populations.
The implications for treatment of individuals with H. pylori
infection were addressed by Nakajima and Hattori . They
systematically reviewed the literature and estimated the expected
annual incidence of esophageal or gastric cancer with and without
eradication of H. pylori infection in patients with chronic
atrophic gastritis. The expected annual incidence of gastric
cancer in patients with corpus atrophy with persistent infection
was at least 5.8-fold higher than that for esophageal
adenocarcinoma after the eradication of infection at all ages.
Even for patients with accompanying reflux esophagitis or
Barrett's esophagus, the incidence of gastric adenocarcinoma with
persistent infection was higher than that of esophageal
adenocarcinoma after eradication of infection. The authors
concluded, therefore, that if eradication of infection lowers the
incidence of gastric cancer, it should be recommended for patients
with corpus atrophy at all ages irrespective of the presence of
reflux esophagitis or Barrett's esophagus, especially in
populations having a high prevalence of gastric cancer .
In summary, increased BMI has been linked with the elimination of
H. pylori infection. As the sphincter mechanism at the
esophagogastric junction is weakened by weight it is not
surprising that obese individuals have a higher incidence of
gastric reflux or GORD . GORD may lead to the development of
Barrett's esophagus, which increases the risk of EA by 40-fold
[132,133]. The study by Lagergren  provides evidence that
these associations may be related as increasing body mass was
associated with a stepwise increase in the risk of EA. If
eradication of H. pylori infection lowers the incidence of gastric
cancer, however, it should be recommended for patients with corpus
atrophy at all ages irrespective of the presence of reflux
esophagitis or Barrett's esophagus, especially in populations
having a high prevalence of gastric cancer .
Attenuated bacteria: Promising carriers of DNA vaccines
Attenuated bacteria will enhance stimulation of the innate immune
system yet increase the safety of a vaccine,  therefore they
may be ideal for the delivery of vaccines. Animal studies have
shown that attenuated S. typhimurium strains can successfully
deliver a variety of genetically engineered DNA vaccine plasmids
for therapeutic vaccination of mice against model tumors
The identification of bacterial "carriers" for DNA vaccines that
target cancer cells by site-specific colonization may allow the
selective delivery of vaccine plasmids into tumor cells .
Colonization of these species may be considered coincidental to
favorable conditions provided by the tumor yet prove clinically
useful. Ultimately, however, the safety and efficacy of
recombinant therapeutic agents expressed by plasmids must be
conducted in appropriate animal models.
Cancer is commonly defined as the uncontrolled growth of abnormal
cells that have accumulated enough DNA damage to be freed from the
normal restraints of the cell cycle. Several pathogenic bacteria,
particularly those that can establish a persistent, infection, can
promote or initiate abnormal cell growth by evading the immune
system or suppressing apoptosis [54,137]. Intracellular pathogens
survive by evading the ability of the host to identify them as
foreign. Other species or their toxins can alter host cell cycles
or stimulate the production of inflammatory substances linked to
DNA damage .
The highly site-specific adherence of bacteria involves binding
species-specific adhesions to the required cell surface receptors.
The role of species that colonize tumors could be causal,
coincidental or potentially protective. If adhesion to the tumor
in question is highly sensitive and specific it may be ideal not
only in diagnosing the presence of a malignancy but also in
delivering the appropriate therapy.
The bacterial species associated with cancer etiology are diverse;
however, the infections they cause share common characteristics
. The time between acquiring the infection and cancer
development is most often years or even decades as seen in cancers
associated with H. pylori, S. typhi and S. bovis infections.
Chronic interactions between the infective agent and immune
response and/or a susceptible host appear to contribute to
carcinogenesis [8,18,38,138]. Preventing or treating the infection
may prevent the cancer in question. Notably, the vast majority of
individuals infected with a cancer-causing species will not
develop cancer .
Evidence suggests that certain individuals are more susceptible to
infections linked to cancer development and that the incidence of
certain cancers varies among populations. For example, gallbladder
cancer is 3 times higher in females as in males in all populations
. Lung cancer is highest in populations that smoke however,
only a small proportion of smokers develop lung cancer .
Although colon cancer is the 3rd highest cancer in the United
States, individuals with IBD have a far greater risk of colorectal
cancer than individuals without IBD [56-58].
A screening test for oral cancer based on salivary counts of
bacterial species is appealing. Currently saliva is meeting the
demand for inexpensive, non-invasive, and easy-to-use diagnostic
aids for oral and systemic diseases, and for assessing risk
behaviors such as tobacco and alcohol use. Although the
colonization of certain bacterial species may be coincidental to
favorable conditions provided by OSCC, increased numbers of
certain salivary species may be clinically useful if shown to be a
signature of oral cancer and if sensitivity and specificity are
Successful treatment for cancers was reported by Dr. Coley and
others one hundred years ago. His approach of using killed
bacterial vaccines was surprisingly effective in some patients
even in the latest stages of cancer. Dr. Coley believed that the
human immune system had the power to cure cancers if properly
stimulated. Today, some investigators agree and have designed new
treatments that stimulate the immune system to recognize and
target the lesion. Recent reports suggest that attenuated
bacterial vaccines can safely and effectively deliver plasmids
encoding tumor self antigens. These studies have reported
successful treatment of certain cancers and prevention of
recurrences [39,110,111]. Cancer vaccines although promising,
present significant challenges. These include identification of
highly effective bacterial strains and their attenuations,
addressing safety issues and the problem of overcoming the
peripheral T cell tolerance against tumor self-antigens .
Further, the response to vaccines will likely vary among
It appears that colonization by certain bacteria may reduce the
risk of cancer in some populations. The epidemiological trends of
esophageal adenocarcinoma and Helicobacter pylori infection have
stimulated research into whether these may be coincidental or due
to an inverse association. Intriguing results suggest there is an
association represented by a complex continuum that begins with
curing infections of virulent strains of H. pylori. The absence of
H. pylori appears to elevate ghrelin which stimulates increased
appetite in some individuals. High ghrelin levels appear to be
associated with increased incidence of obesity. Obesity is
reported to be a contributing factor in GORD. Finally, GORD may
lead to Barrett's esophagus which increases the risk of esophageal
adenocarcinoma. If these relationships can be proven, then the
colonization of this species and its seemingly negative
association with EA may be more clearly understood.
In summary, recent research has uncovered a great deal of
information regarding the bacterial mechanisms used to cause,
colonize or cure cancer, however, many questions remain. For
example, do the bacteria in question initiate, promote, or merely
show affinity for the neoplasm? Conversely does cancer weaken the
host which facilitates acquiring the infection? Can the highly
site specific colonization of certain bacteria for a tumor be
clinically useful in diagnosis or treatment? Could attenuated
bacteria be used in vaccines to safely and effectively deliver
therapeutic agents? The continued exploration of these questions
will bring research ever closer to the prevention, early diagnosis
and truly effective treatment of this scourge of mankind.